DE69713285T2 - 17-BETA-CYCLOPROPYL (AMINO / OXY) 4-AZA STEROIDS AS TESTOSTERONE 5-ALPHA-RECTASE AND C17-20-LYASE INHIBITING COMPOUNDS - Google Patents

Abstract

The invention related to 4-aza-17 beta -(cyclopropoxy)-androst-5 alpha -androstan-3-one, 4-aza-17 beta -(cyclopropylamino)-androst-4-en-3-one and related compounds and to compositions incorporating these compounds, as well as the inhibition of C17-20 lyase, 5 alpha -reductase and C17 alpha -hydroxylase and to the use of these compounds in the treatment of androgen and estrogen mediated disorders, including benign prostatic hyperplasia, androgen mediated prostate cancer, estrogen mediated breast cancer and to DHT-mediated disorders such as acne. Disorders relating to the oversynthesis of cortisol, for example, Cushing's Syndrome, are also included. The treatment of androgen-dependent disorders also includes a combination therapy with known androgen-receptor antagonists, such as flutamide. The compounds of the invention have general formula (I).

Description

BACKGROUND OF THE INVENTION

The enzyme steroid C₁₇₋₂₀-lyase cleaves the 17-20 carbon-carbon bond in steroids having a two-carbon side chain at the position of the 17β carbon atom, thereby forming important precursor molecules for the formation of testosterone, 5α-dihydrotestosterone and the estrogens, primarily estrone and estradiol. Compounds that inhibit this enzyme would thus serve to inhibit the formation of the indicated precursors and thereby be useful in the treatment of various androgenic as well as estrogenic disorders. Treatment involving such enzyme inhibitors is not limited to the formation of the precursor molecule, as are various organ removal procedures currently known. For example, although a testicular removal will effectively reduce gonadal androgen, it will have no significant effect on adrenal androgen production. In addition, such enzyme treatment is a much more focused treatment in that it is directed at the immediate hormonal imbalance believed to be responsible for the condition, as opposed to a broad-spectrum remedy that will not only affect the particular symptom but will cause permanent endocrine damage requiring a lifelong dependence on replacement therapy.

Furthermore, certain types of breast cancer are known to be estrogen dependent. Adrenalectomy, ovarianectomy, and pituitaryectomy have been used, as well as non-surgical procedures that result in tumor regression. Human patients with advanced breast cancer who are given estrogen biosynthesis enzyme inhibitors have been shown to experience dramatically reduced plasma estradiol levels and improved therapeutic effects, at least as effective as adrenalectomy. (Jean Van Wauve and Paul A. J. Janssen, Journal of Medicinal Chemistry, 32, 10: 2231-2239).

Prostate cancer, or neoplastic tissue disorders originating in the parenchymal epithelium of the prostate, are among the most common malignancies in men and have the highest cancer-specific death rate of all malignant carcinomas. Patients with metastatic prostate cancer are known to respond positively to hormone therapy. Cookson and Sarosdy report that androgen removal has a positive, favorable response for 60% to 80% of all patients studied. (CS Cookson and MF Sarosdy, South Med. J. 87: 1-6).

More specifically, C17,20-lyase inhibitors would be useful in the treatment of hormone-dependent prostate carcinoma, prostatic hyperplasia, virilism, congenital adrenal hyperplasia due to 21-hydroxylase deficiency, hirsutism, hormone-dependent breast cancer, polycystic ovary syndrome associated with increased C17,20-lyase activity, and other neoplastic tissue disorders, such as endometrial, hepatocellular and adrenal carcinomas.

The enzyme steroid 5α-reductase, present in mammalian tissues including skin, male sex organs and the prostate, catalyzes the conversion of testosterone (17β-hydroxyandrostan-4-en-3-one) to dihydrotestosterone or DHT (17β-hydroxy-5α-androst-3-one), also known as stanolone. DHT is a more potent androgen than testosterone, and functions as a sensory end-organ effector in certain tissues, particularly in mediating growth. DHT formation may occur in certain tissues by the action of 5α-reductase. In the treatment of androgen-dependent disorders such as benign prostatic hyperplasia and prostate cancer, including hormone-dependent carcinomas, inhibition of DHT would be highly desirable.

The conversion of testosterone to DHT itself can be associated with various androgenic disorders, particularly when DHT levels build up to excessive amounts. For example, high DHT levels in the skin have been linked to the pathogenesis of acne, including acne vulgaris.

Agents having the ability to inhibit both C17-20 lyase and 5α-reductase would inhibit not only DHT production but also testosterone formation. By inhibiting the major androgenic steroid hormones, such compounds would have enhanced utility in treating androgenic disorders. Such dual inhibitors with a 17-cyclopropyl substitution but without the 4-aza nitrogen atom are disclosed in WO-A-94/28010.

The enzyme C₁₇-hydroxylase catalyzes the C₁₇-hydration of steroid substrates during the biosynthesis of cortisol. Since C₁₇-20 lyase and C₁₇-hydroxylase are different sites of action of the same enzyme, inhibition of one normally leads to inactivation of the other. Cortisol excess leads to a syndrome characterized by hypokalemia, metabolic alkalosis, polydipsia, polyuria, Cushing's syndrome and hypertensive complaints. Inhibition of cortisol synthesis via C₁₇α-hydroxylase would therefore have a therapeutic effect for the treatment of these disorders or complaints.

SUMMARY OF THE INVENTION

The present invention relates to 4-aza-17-(cyclopropoxy)androst-5α-androstan-3-one, 4-aza-17-(cyclopropylamino)androst-4-en-3-one and related compounds and compositions including these compounds, as well as the use of these compounds in the treatment of conditions that would be affected by inhibition of C17-20 lyase and/or 5α-reductase, including androgen and estrogen mediated disorders such as benign prostatic hyperplasia, DHT mediated disorders such as acne, estrogen dependent breast cancer and androgen mediated prostate cancer. Since the present compounds also inactivate the action of C₁₇α-hydroxylase, disorders characterized by excessive synthesis of cortisol can also be treated by the compounds of the invention. For example, hypokalemia, metabolic alkalosis, polydipsia, polyuria, Cushing's syndrome and hypertensive disorders.

In another embodiment, the compounds of the invention can be administered together with another effective treatment for enhanced therapeutic effect. For example, in the treatment of androgen dependent disorders, including prostate cancer, flutamide, a known androgen receptor antagonist, can be used together with the compounds of the invention.

More specifically, the present invention is directed to a group of compounds and their pharmaceutically acceptable salts having the following general formula:

in the:

A means O or NH;

R¹ represents H or C₁₋₄alkyl;

R² represents H, a halogen atom, a phenylthio, phenylsulfinyl or phenylsulfonyl group ;

R³ represents H, a halogen atom, a C₁₋₄alkylthio, C₁₋₄alkylsulfinyl or C₁₋₄alkylsulfonyl radical;

R⁴ is H, C₁₋₄ alkyl or C₂₋₄ alkenyl;

R⁵ represents H or C₁₋₄ alkyl;

A:

a) an oxo group;

b) (H)(H) or an α-hydrogen atom and a β-substituent selected from a C₁₋₄alkyl, C₂₋₄alkenyl group, a hydroxy group, a C₁₋₄alkanoyloxy group, C₁₋₄alkoxycarbonylethyl group, a carboxymethyl group, a C₁₋₄alkoxycarbonyl group, a carboxy group, a C₁₋₄alkanoyl group and a halogen atom;

provided that:

a) a 1,2-double bond is present when R² is present and is different from a hydrogen atom,

b) no 6,7-double bond is present when Z is an oxo group, and

c) A does not represent an NH group when R¹ represents a CH₃ group and R², R³, R⁴ and R⁵ represent H and Z represents(H)(H) and there are no double bonds in any of the positions 1(2), 5(6) or 6(7).

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the term "C1-4 alkyl" means any linear or branched chain alkyl radical having one to four carbon atoms. For example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, and the like.

As used herein, the term "C2-4 alkenyl" means any linear or branched chain alkene radical of two to four carbon atoms. For example, ethenyl, vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1,3-butenyl, and the like.

As used herein, the term "C1-4 alkylthio, C1-4 alkylsulfinyl or C1-4 alkylsulfonyl" means a C1-4 alkyl-Y- group, where the C1-4 alkyl group is as defined above and Y is an S-, SO- or SO2 radical, respectively, and as illustrated in Scheme G. "Phenylthio, phenylsulfinyl or phenylsulfonyl group" is defined in a similar manner or Ph-S-, Ph-SO- or Ph-SO2-.

As used herein, the term "C1-4 alkanoyloxy" defines a final product molecule which is the ester condensation product of the corresponding steroid alcohol with a linear or branched chain carboxylic acid having one to four carbon atoms. For example, formyloxy, acetyloxy, n-propionyloxy, isopropionyloxy, n-butanoyloxy, s-butanoyloxy, t-butanoyloxy group, and the like. It is graphically represented by compound 49 in Scheme I or compound 56 of Scheme J.

As used herein, the term "C1-4 alkoxycarbonylmethyl" means a C1-4 alkyl (as defined above) ester of acetic acid, all of which form a substituent attached to the α-carbonyl carbon atom of the steroid nucleus as shown in Scheme K.

As used herein, the term "C1-4 alkoxycarbonyl" means a C1-4 alkyl (as defined above) ester of formic acid, all of which form a substituent attached to the carbonyl carbon atom of the steroid nucleus as shown in Scheme L.

As used herein, the term "C1-4 alkanoyl" means a ketone of one to four carbon atoms attached to the steroid nucleus at the carbonyl carbon atom as shown in Scheme M. For example, an ethanoyl, isopropanoyl, n-butanoyl, s-butanoyl, t-butanoyl group.

As used herein, the term "halogen atom" means a chlorine, bromine, fluorine or iodine substituent.

As used herein, the term "pharmaceutically acceptable salt" is intended to mean any organic or inorganic acid salt capable of forming a nontoxic acid addition salt suitable for use as a medicament. Illustrative inorganic acids that form suitable salts include hydrochloric, hydrobromic, sulfuric and phosphoric acids and acidic metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids that form suitable salts include the mono-, di- and tricarboxylic acids. For example, acetic, glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, hydroxybenzoic, phenylacetic, cinnamic, salicylic, glutamic, gluconic, formic and sulfonic acids such as methanesulfonic acid and 2-hydroxyethanesulfonic acid. Further examples of suitable pharmaceutically acceptable salts are listed by S. M. Berge et al. J. Pharm Sci. 66: 1, 1 (1977), which is incorporated herein by reference. Such salts may be in either hydrated or substantially anhydrous form.

As used here, the term "patient" refers to a warm-blooded animal, such as a mammal, suffering from a particular disease. It is explicitly understood that guinea pigs, dogs, cats, rats, mice, horses, cattle, sheep, and humans are examples of animals within the meaning of the term.

As used herein, the term "effective inhibitory amount" is such an amount at which an enzyme inhibitory effect is achieved which is sufficient to cause a therapeutic effect in the patient. The exact amount of the compound to be administered can be readily determined by the attending diagnostician, as one of skill in the art, by using conventional procedures and by observing results obtained under analogous circumstances. Factors significant in determining the dose include: the dose; the type of animal, its size, age and general state of health; the particular disease involved, the extent or complexity or severity of the disease; the response of the individual patient; the particular compound administered; the mode of administration; the bioavailability characteristics of the preparation administered; the dosage regimen selected; the use of concomitant medication, and other relevant circumstances. That is, the exact amount used can vary over a wide range. For example, from about 0.625 to 200 mg/kg body weight per day, preferably from about 0.5 mg to 100 mg/kg body weight per day.

In practicing the methods of this invention, the active ingredient is preferably incorporated into a composition containing a pharmaceutical carrier, although the compounds are effective and can be administered alone. The term "pharmaceutical carrier" refers to known pharmaceutical excipients that are useful in formulating pharmaceutically active compounds for administration and that are substantially nontoxic and nonsensitizing under the conditions of use. The precise proportion of these excipients will be determined by the solubility and chemical properties of the active compound, the route of administration chosen, and standard pharmaceutical practice. That is, the proportion of active ingredient may vary from about 5% to about 90% by weight.

FORMULATIONS:

The pharmaceutical compositions of the invention are prepared in a manner well known in the pharmaceutical art. The carrier or excipient may be a solid, semi-solid or liquid material which can act as a vehicle or medium for the active ingredient. Suitable carriers or excipients are well known in the art. The pharmaceutical composition may be adapted for oral, inhalation, parenteral or topical use and may be administered to the patient in the form of tablets, capsules, aerosols, inhalants, suppositories, solutions, suspensions, powders, syrups and the like. As used herein, the term "pharmaceutical carrier" means one or more excipients.

In preparing formulations of the compounds of the invention, efforts should be made to ensure bioavailability of an effective inhibitory amount, including oral, parenteral and subcutaneous routes. For example, effective routes of administration may include subcutaneous, intramuscular, intravenous, transdermal, intranasal, rectal and the like, including delivery from implants as well as direct injection of the active ingredient and/or composition directly into the tissue or tumor sites. Suitable pharmaceutical carriers and formulation methods are found in standard texts such as Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania, which is incorporated herein by reference.

For oral administration, the compounds may be formulated into a solid or liquid preparation such as capsules, pills, tablets, lozenges, powders, solutions, suspensions or emulsions, with or without inert diluents or edible carriers. The tablets, pills, capsules, lozenges and the like may also contain one or more of the following adjuvants: binding agents such as microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, Primogel®, corn starch and the like; lubricants such as stearic acid, magnesium stearate or Sterotex®; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; and flavoring agents such as peppermint, methyl salicylate or fruit flavor. If the unit dosage form is a capsule, it may also contain a liquid carrier such as polyethylene glycol or a fixed oil. The materials used should be pharmaceutically pure and nontoxic in the amounts used.

For parenteral administration, the compounds may be administered as injectable dosages of a solution or suspension of the compound in a physiologically acceptable diluent with a pharmaceutical carrier, which may be a sterile liquid such as water-in-oil, with or without additions of a surfactant and other pharmaceutically acceptable excipients. Illustrative of oils that may be used in the preparations are those of petroleum, animal, vegetable or synthetic origin. For example, peanut oil, soybean oil and mineral oil. In general, water, saline, aqueous dextrose and related sugar solutions, ethanols and glycols such as propylene glycol are preferred liquid carriers, particularly for injectable solutions. The parenteral preparation may be enclosed in ampoules, disposable syringes or multi-dose vials made of inert glass or plastic.

The solutions or suspensions described above may also include one or more of the following adjuvants: sterile diluents such as water for injection, saline, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as ascorbic acid or sodium bisulfite; Complexing agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates; and tonicity adjusters such as sodium chloride or dextrose.

The compounds may be administered in the form of a skin patch, depot injection or implant preparation which may be formulated in such a manner as to allow sustained release of the active ingredient. The active ingredient may be compressed into pellets or small cylinders and implanted subcutaneously or intramuscularly as depot injections or implants. Implants may use inert materials such as biodegradable polymers and synthetic silicones. Further information on suitable pharmaceutical carriers and formulation procedures can be found in standard texts such as Remington's Pharmaceutical Sciences.

CHEMICAL SYNTHESIS

The following reaction schemes and descriptive text describe the preparation of the various compounds of the invention. Various combinations and permutations to achieve individual compounds will be immediately apparent to one of ordinary skill in the art.

Scheme A represents one possible synthesis for the C17-cyclopropyl-5-ene steroid compounds of the invention starting from testosterone. Testosterone or 17β-hydroxyandrost-5(6)-en-3-one [1] is treated with a strong oxidizing agent which disrupts the A-ring of the steroid nucleus to give the corresponding 4-nor-3,5-seco-acid [2]. For example, potassium permanganate with sodium periodate in aqueous potassium carbonate and tert-butanol or methanolic ozone in methylene chloride at reduced temperature have been shown to be effective. However, care should be taken to ensure that overoxidation does not occur, thereby converting the C17-hydroxy substituent to a ketone.

The seco acid [2] can be converted to the corresponding lactam or 4-azasteroid [3] by refluxing it in the presence of an acidic ammonium addition salt and an acid. For example, ammonium acetate in acetic acid. The corresponding 4-alkylaza compounds of the invention can be prepared by refluxing with the appropriate alkylamine or alkylamine hydrochloride under acidic conditions. For example, to prepare the desired 4-methyl-4-azasteroid, the seco acid [2] is refluxed with methylamine hydrochloride in the presence of acetic acid. The acid addition ester [3] can be converted to the corresponding 17-alcohol [4] under basic hydrolysis conditions, such as aqueous sodium hydroxide in ethanol. Tetrahydrofuran (THF) may be used if necessary to aid the solubility of the steroid substrate.

The 17-alcohol [4] can be converted to the vinyl ether [5] by etherification with a vinyl ether in the presence of a suitable etherification catalyst and solvent. For example, ethyl vinyl ether and mercury(11) acetate in chloroform or chloroform/tert-butyl methyl ether. AB Charette et al., Tet. Lett. 35(4), 513-516 (1994). The vinyl ether [5] can then be converted to the cyclopropyl ether [6] under typical cyclopropanation conditions, such as by reaction with tert-butyl methyl ether, diethylzinc and methylene iodide in methylene chloride.

Scheme B graphically illustrates a synthetic route for preparing the saturated B-ring C17 cyclopropyl ether compounds of the invention. In Scheme B, Option A, the 5-ene-C17 acid ester [3] is hydrogenated to the saturated acid ester [7] and then hydrolyzed to the saturated 17-alcohol [8]. Typical hydrogenation conditions include heating with hydrogen in the presence of ethanol and 5% palladium on carbon catalyst. The hydrolysis conditions are similar to those described under Scheme A, aqueous sodium hydroxide in ethanol and tetrahydrofuran, with the solvent being chosen as necessary to dissolve the reactants. However, the hydrogenation and hydrolysis steps can be interchanged, i.e., in option B, the 5(6)-unsaturated 17-alcohol [4] is prepared directly by hydrolyzing the acid ester [3] and then hydrogenated to give the saturated 17-alcohol [8]. The 17-alcohol [8] can then be etherified and cyclopropanated as described in Scheme A to give the 17β-cyclopropyl ether [9]. By "inert substituent" in the definition of R' is meant substituent(s) that are not affected by the reaction conditions of the scheme.

Scheme C illustrates one possible synthesis for preparing the compounds of the invention having a 17-cyclopropylamino substituent. The starting compound testosterone [1] is treated under oxidation conditions sufficient to cleave the A ring of the steroid nucleus to give the corresponding 17-keto-4-nor-3,5-seco acid [10]. This can be carried out in a manner similar to that described for preparing compound [2] in Scheme A. Since oxidation of the C17 hydroxy substituent to the C17 ketone is desirable, reaction conditions modified from the ring cleavage of Scheme A are preferably used. For example, bubbling ozone at reduced temperature (-78°C) into methylene chloride and ethyl acetate. Non-alcoholic solvents are used to ensure that no transesterification with the newly formed seco acid occurs. The seco acid [10] is converted to the corresponding 17-ketolactam or 4-azasteroid [11] using conditions similar to those described for the corresponding reaction in Scheme A. For example, heating to reflux in the presence of ammonium acetate and acetic acid. The 4-alkyl compounds can be prepared in a similar manner, e.g., by heating to reflux in acidic alkylamine or acidic alkylamine hydrochloride. To obtain the 5(6)-olefin as defined by Route A, the 17-ketolactam [11] is converted to the corresponding 17-cyclopropylimino compound [12] by reaction with cyclopropylamine in chloroform. THF can be used as a cosolvent if necessary to solubilize the steroid substrate. The cyclopropylimine [12] is reduced to the corresponding 17-cyclopropylamine [13] by reaction with a suitable reducing agent, such as sodium borohydride.

The saturated cyclopropylamino compounds of the invention can also be prepared under Scheme C following Route B. The 17-ketolactam [11] is preferably hydrogenated by the action of H2 gas with palladium catalyst to give the saturated 17-hydroxylactam [14]. The 17-alcohol [14] can be oxidized to the corresponding 17-ketone [15] by reaction with tetrapropylammonium perruthenate (VII) (TPAP) and 4-methylmorpholine N-oxide (NMO) in the presence of 4 Å molecular sieves. The 17-ketone [15] is then converted to the corresponding cyclopropylimine [12] and reduced to give the 17-cyclopropylamino compound [13] in a similar manner as described in Route A above.

Scheme D illustrates a possible synthesis for the 1-halo-Δ1 compounds of the invention. The synthesis can start with the saturated acid ester [7], also an intermediate of Scheme B. To obtain the 1-halo-Δ1-4-aza compounds of the invention, the acid ester [7] is preferably dehydrogenated at the Δ1(2) positions to give the corresponding 1(2)-ene [16] as is known. For example, reaction with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) and bis(trimethylsilyl)trifluoroacetamide (BSTFA) in dioxane. Bhattacharya et al., J. Am. Chem. Soc., 1988, 110, 3318-3319. In Option A, the hydrolysis, vinylation and cyclopropanation can be carried out in a manner similar to that described under Scheme A to give the Δ1(2)-ene-17-cyclopropyl ether [17]. In Option B, the 17-cyclopropylamine [17] is formed by first hydrolyzing to the 17-alcohol as in Option A, but then oxidizing, cycloaminating and reducing as described under Route B in Scheme C.

Compound [17] can be converted into the corresponding 1-phenylthioether [18] by reaction with phenyl mercaptan (thiophenol) in sodium hydride. The thioether [18] can be converted into the 1,2-dihalo compound [19] by reaction with N-bromosuccinimide (NBS) and diethylaminosulfur trifluoride (DAST). R. Bohlmann, Tetrahedron Lett. 1994, 35(I), 85-88. The 2-halo substituent can then be eliminated by reaction with tributyltin hydride and azobisisobutyronitrile to give the desired 1-fluoro-Δ1(2) compound [20]. This compound [20] can, if desired, be hydrogenated by reaction with H₂ gas over palladium to give the saturated 1-fluoro compound [21].

Scheme E graphically represents a possible synthesis for the 1-phenylsulfinyl and 1-phenylsulfonyl compounds of the invention starting from the 1-phenylthioether [18], the preparation of which is described in Scheme D, as is known in the art. For example, compound [18] can be reacted with 3-chloroperoxybenzoic acid at reduced temperature (-78°C) for 3 hours under nitrogen to produce the 1-phenylsulfinyl thioether. The 1-phenylsulfonyl thioether is prepared under similar reaction conditions as the sulfinyl ether, except that the reaction occurs at room temperature and the time is extended to 16 hours.

Scheme F represents the preparation of the 2α-halo compounds of the invention starting from the 4-aza-17-alcohol [14], the preparation of which is described in Scheme C, Route B. The 17-alcohol [14] is first converted to the protected ether [24] by any effective means, for example by reaction with trimethylsilyl chloride in methylene chloride. The protected ether [24] can then be halogenated by reaction with N,N,N',N'-tetramethylethylenediamine (TMEDA) and the desired halogenated silyl agent at reduced temperature under an inert atmosphere. Once halogenated, the protecting group is removed for subsequent conversion of the 17-substituent. For example, to prepare the bromide [25], trimethylsilyl iodide and bromine in TMEDA and toluene can be used initially, followed by tetrabutylammonium fluoride (TBAF) in tetrahydrofuran (THF). Accordingly, the iodide [26] can be prepared by trimethylsilyl chloride and iodine in TMEDA and toluene, followed by the action of TBAF in THF. The 2α-halogen compounds [25] and [26] can then be converted into the corresponding 17β-cyclopropylamino compounds [27] and [28] as in Scheme C or the corresponding 17β-cyclopropyloxy compounds [27] and [28] as described in Scheme A.

Scheme G represents a synthesis for preparing the 2α-alkylthio/alkylsulfinyl and alkylsulfonyl compounds of the invention starting from the 2α-iodocyclopropyl ether or cyclopropylamino compound. [28] The alkyl thioether [29] can be prepared by reaction with the corresponding alkali metal salt of the alkylthiol in a suitable solvent as is known. For example, the methyl thioether can be prepared by using sodium thiomethoxide (sodium methyl sulfide; sodium methanethiolate) in ethanol. The sulfoxide [30] and sulfone [31] can then be prepared as in Scheme E.

Scheme H1; represents a synthesis for generating the 7β-alkyl compounds of the invention starting from 17β-hydroxyandrost-5-en-3-ol-3-acetate [32]. In Scheme H1, compound 32 is first protected with any suitable protecting agent. For example, t-butyldimethylsilyl chloride and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) in methylene chloride. The protected acetate [33] is then C7-oxidized to produce the 7-ketone [34] by any known means. For example, by reaction with t-butyl chromate in acetic anhydride, acetic acid and carbon tetrachloride. A. Pinto et al., Chem. Pharm. Bull. 1988, 36(12), 4689-4692. The 7-ketone is then reacted with the appropriate Grignard reagent to give the corresponding 7-alkyl-7-alcohol [35]. For example, the 7-ethyl-7-alcohol can be formed by reaction with ethylmagnesium chloride in THF. For example, the 7-aryl-7-alcohol can be formed by reaction with 4-bromotolylmagnesium chloride in THF.

The 7-substituted 7-alcohol [35] is dehydrogenated to the 7-alkyldiene [36] by reaction in a suitable material, such as by reaction with aluminum isopropoxide in the presence of toluene and cyclohexanone. JF Eastham & R. Teranihi, Org. Synth. Coll. Vol. IV 1963, 192-195; C. Djerassi, Org. React. 1951, 6, 207-272. The diene [36] can then be hydrogenated and isomerized under known conditions to give the olefin [37]. For example, reaction with dry ammonia and lithium metal in t-butanol and toluene. Crabtree et al., Org. Synth. 1991, 70, 256-264; D. Caine et al., Org. Synth. Coll. Vol. VI 1988, 51-55; D. Caine, Org. React., 1976, 23, 1-258.

In Scheme H2, the olefin [37] can be isomerized and desilylated to the 4-ene-17-alcohol [38] by any suitable reagent. For example, treatment with 1,8-diazabicyclo[5.4.0]undec-7-ene at reflux, followed by cooling to room temperature and treatment with tetrabutylammonium fluoride. Under route A, compound [38] is then oxidized, lactamized and hydrolyzed as described in Scheme A to give the 4-aza-17-alcohol [39], which can then be etherified and cyclopropanated as in Scheme A to give the corresponding cyclopropyl ether [40].

In the generation of cyclopropylamine [42] using route B, the 17-alcohol [39] can be oxidized to the ketone [41] and then cycloaminated and reduced as in Scheme C, route B However, it is also immediately apparent that the cyclopropylamine can be prepared from the 17-alcohol [38] with fewer synthetic steps by performing the oxidative ring cleavage, lactamization, cycloamination and reduction as described in Scheme C, Path A (not shown in Scheme H2).

Scheme I briefly describes a synthesis for the 7-hydroxy-, 7-oxo- and 7-alkanoyloxy-17β-cyclopropyl ether compounds of the invention from 3-β-acetoxy-17β-t-butyldimethylsilyloxyandrost-5-en-7-one [34]. Compound [34] is ketalized under suitable known reaction conditions, such as reaction with 1,2-bis(trimethylsiloxy)ethane and trimethylsilyl trifluoromethanesulfonate in methylene chloride at -78°C, to give the 7-ketal-3-acetate [43]. T. Tsunoda et al., Tetrahedron Lett, 1980, 21 (14), 1357-1358; JR Hwu et al., J. Org. Chem. 1987, 52(2), 188-191. Compound [43] is hydrolyzed as in Scheme A, followed by oxidation to the 3-ketone [44]. The oxidation can be carried out, for example, similarly to the conversion of compound [35] to [36] in Scheme H1, i.e., heating to reflux with aluminum isopropoxide in the presence of toluene and cyclohexanone. The 3-ketone [44] is then desilylated/isomerized, oxidized, lactamized, and hydrolyzed as described in Scheme H2 to afford the 17-alcohol [45]. The 17-alcohol [45] can then be etherified and cyclopropanated as in Scheme A to afford the 3,7-dioxo-17β-cyclopropyl ether [46].

The cyclopropyl ether [46] can be directly reduced to the 7-alcohol [48] as indicated in option A, or it can be first hydrogenated to compound [47] and then reduced as indicated in option B. The reduction conditions can be similar to those used in previous schemes, for example, sodium borohydride in ethanol and THF. The hydrogenation can also be carried out in a manner similar to that described above (Scheme B), for example, heating in the presence of hydrogen and palladium catalyst.

The cyclopropyl ether-7-alcohol [48] can be esterified to alkyl alcohol esters [49] by reaction with the appropriate alkyl anhydrides. For example, to prepare the alcohol ester of propionic acid, compound [48] is reacted with propionic anhydride in pyridine. HH Baer et al., Can. J. Chem. 1991, 69, 1563-1574.

Scheme J illustrates the preparation of the 7-oxo-, 7-hydroxy- and 7-alkoxycarbonyl-17-cyclopropylamino compounds of the invention starting from 17β-hydroxy-3,7-dione [45], also an intermediate of Scheme I. Compound [45] is first deprotonated and then immediately silylated to produce the protected ether [50]. Suitable conditions include, for example, reaction with lithium diisopropylamide followed by trimethylsilyl chloride at -78°C. The protected ether [50] can then be dehydrated and 7-silylated and acid hydrolyzed in the usual manner. Suitable reaction conditions include, for example, treatment with lithium diisopropylamide in THF at -78°C followed by addition of t-butyldimethylsilyl chloride. Once this reaction product is worked up, it can be acidic hydrolyzed with acetic acid in THF to give the 7-protected 17-alcohol ether [51]. Compound [51] can then be oxidized, cycloaminated and reduced as in Scheme C, Path B to give the protected cyclopropylamine [52].

Compound [52], when deprotected, affords the 3,7-dioxocyclopropylamine [53]. Suitable conditions for this transformation include, for example, tetrabutylammonium fluoride in THF under an inert atmosphere. The remaining compounds [54], [55] and [56] in the scheme can be prepared in a manner similar to that described under Scheme I.

Scheme K graphically illustrates one possible preparation of the 7-alkoxycarbonylmethyl and 7-carboxymethyl compounds of the invention starting from compound [50], also an intermediate in Scheme J. The protected 7-ketone [50] is carboxylated to form the illustrated alkylcarbonyloxymethyldiene [57]. For example, the 7-ethylmethylcarboxylate can be prepared by reaction with triethylphosphonoacetate in THF and sodium hydride. The diene [57] can then be desilylated and hydrogenated to the 17-alcohol [58] in a typical manner, such as by treatment with tetrabutylammonium fluoride in THF. The 17-alcohol [58] can be converted to the cyclopropyl ether [59] prepared under Option A, similarly as described in Scheme A. Alternatively, the 17-alcohol [58] can be converted to the cyclopropylamine [59] prepared under Option B, similarly as described in Scheme C. Compound [59] can then be hydrolyzed in a conventional manner (Scheme A) to give 7-ethanoic acid [60].

Scheme L graphically illustrates a synthesis of the 7-alkoxycarbonyl and 7-carboxylic acid compounds of the present invention starting from the 7-alkyl ethanoate [58], also an intermediate in Scheme K. The carbonyl group is phenylated in a conventional manner, for example by reaction with phenylmagnesium chloride (4 mol eq.) in THF, to give the 7-diphenylmethyl alcohol [61]. This compound [61] can then be dealkylated to the 7-acid [62] as is known. For example, reaction with chromium trioxide (chromic acid) in water, methylene chloride and acetic acid. B. Riegel et al., Org. Synth. Coll. Vol. 3, 1955, 234-236; CS Subramaniam et al., Synthesis 1978, 468-469. The 7-acid [62] can then be converted to the alkyl ester [63] as is known. For example, 4-(dimethylamino)pyridine and 1,3-dicyclohexylcarbodiimide in methylene chloride and ethanol. B. Neises and W. Steglich, Org. Synth. 1984, 63, 183-187. The ester [63] can then be converted to either the cyclopropyl ether [64A] (Scheme A) or the cyclopropylamine [64B] (Scheme C) as described above. Compound [64] can then be hydrolyzed in a conventional manner (e.g. NaOH in water, ethanol and/or THF) to the 7-acid [65].

Scheme M illustrates the preparation of the 7α-ketone compounds of the invention starting from the 7-alkyl ester [63]. The ketone [63] is first reduced to the corresponding alcohol in a conventional manner (e.g. sodium hydride in ethanol) and silylated as in Scheme H1 to give the protected ester [66]. The ester [66] is then reduced to the 7-methyl alcohol [67]. Suitable reduction conditions include, for example, lithium borohydride in THF. RW Jeanloz & L. Walker, Carbohydrate Res. 1967, 4, 504 and ERH Walker, Chem. Soc. Rev. 1976, 5, 23-50. The alcohol is then oxidized to the 7-aldehyde [68]. Suitable oxidation conditions include, for example, 4-hydroxy-TEMPO-benzoate in methylene chloride and sodium bicarbonate followed by sodium bromite. T. Inokuchi et al., J. Org. Chem. 1990, 55, 462-466. The 7-aldehyde [68] is then alkylated, then oxidized and desilylated in the usual manner (Schemes C and J, respectively) to give the 17-hydroxy-7-alkanone [69]. For example, to prepare the 1-propanone, titanium(IV) chloride and tetraethyl lead are added sequentially at -78 °C in methylene chloride. T. Yamamoto and JI Tamada, J. Am. Chem. Soc. 1987, 109, 4395-4396. Compound [70] can then be prepared as a cyclopropyl ether (choice A, Scheme A) or as a cyclopropylamine [70]. In preparing the cyclopropylamine, the 7-alkanoyl ester must first be protected by suitable means such as by formation of the ethylene or 2,2-dimethylpropane ketal, followed by the steps described in Option B, Scheme C, and subsequent deprotection. For example, deprotection can be carried out by ethylene glycol or 2,2-dimethylpropane-1,3-diol with acid catalysis, respectively, and the deprotection is carried out under acidic conditions, taking care to minimize reaction with the acid-sensitive C17 cyclopropylamine, as is known.

Scheme N graphically illustrates a possible synthesis for the C16 alkenyl and C16 alkyl compounds of the invention starting from androstenedione (androst-4-ene-3,17-dione) [72]. Compound [72] is treated in a manner similar to the procedure described in Scheme C to prepare the azaandrostenedione [11]. The dione can then be C16 alkylated by known methods to give the 16-alkenyldione [73]. For example, to prepare the 16α-allyldione, diethyl oxalate and sodium methoxide are added sequentially in methylene chloride solvent at 0°C, followed by reaction with methyl iodide at 55°C and finally treatment with sodium methoxide. NI Carruthers et al., J. Org. Chem. 1992, 57 (3), 961-965. The alkenyldione [73] can then be converted to the cyclopropyl ether (Scheme A) or to the cyclopropylamine (Scheme C) as described above to give the 16-alkene [74]. The 16-alkene [74] can then be hydrogenated in the usual manner [Scheme B] to give the 16-alkane [75].

Scheme O1 graphically represents the first part of a synthesis for preparing the 15-alkyl compounds of the invention starting from dehydroisoandrosterone 3-acetate (3β-acetoxy-5-androsten-17-one). The 3-acetoxy-17-one [76] can be ketalized at the C17 position in the usual manner (Scheme I) to give the 17-ketal [77]. The ketal [77] is α-brominated to give the 16-bromide [78]. Suitable bromination conditions include, for example, pyridinium bromide perbromide in dry THF followed by treatment with sodium iodide then reaction with sodium thiosulfate in water and pyridine. The bromide [78] can then be dehydrogenated and 17-hydrolyzed to the 15-en-17-one. Typical dehydrogenation conditions include, for example, potassium t-butoxide in dimethyl sulfoxide. Typical hydrolysis conditions include, for example, p-toluenesulfonic acid monohydrate. The ketone produced by hydrolyzing the ketal can then be silylated in a conventional manner (Scheme H1) to give the silylated diene [79]. The silylated diene [79] can then be selectively alkylated at C15 to give the 15-alkylsilylated 17-ketone [80], as is known in the art. For example, to prepare the 15-ethyl compound, compound [79] can be added dropwise to ethylmagnesium chloride in ether which has been previously treated with copper(L) chloride in THF. The silylated ketone [80] can then be deprotected and oxidized in a conventional manner (Scheme I or Scheme H1 [35] to [36]) to give the alkylated dione [81]. The alkylated dione [81] is then converted to the seco acid (ring opening) and ring closure in a typical manner as described in Scheme C leads to the azadione [82].

Scheme O₂ represents the second part of the synthesis of the 15-alkyl compounds of the invention. The azadione [82] can be converted directly (path B) to the desired cyclopropyl ether [84A] (option A) or cyclopropylamine [84B] (option B) in a conventional manner (Scheme A or Scheme C). Alternatively, the azadione (path A) can be hydrogenated under typical conditions (Scheme N) to give the 15-alkylazaandrostane [83], which can be converted to either the cyclopropyl ether [84A] or the cyclopropylamine [84B] as described above.

7-alkyl compounds

The 7-alkyl compounds of the invention can be prepared in a manner analogous to that described in PCT applications PCT/US/04643 (WO 93/23420) and PCT/US/04734 (WO 93/23039), the disclosures of which are incorporated herein by reference.

7-Alkenyl, carboxy and methylcarboxy compounds

The compounds of the invention having an alkenyl, carboxy or methylcarboxy substituent in the 7-position can be prepared in a manner analogous to that presented in PCT applications PCT/US/04643 (WO 93/23420) and PCT/US/04734 (WO 93/23039), the disclosures of which are incorporated herein by reference.

2-halogenated compounds

The compounds of the invention having a halogen substituent in the 2-position can be prepared by the process described in European Patent Application 0473225 A2 (91-202135), the disclosure of which is incorporated herein by reference.

2-Halogen, R-Thio, R-Sulfinyl, R-Sulfonyl compounds

The compounds of the invention in which the above 2-substituents are present can be prepared in a manner analogous to that described in European Patent Application 0473226 A2 (91-202135), the disclosure of which is incorporated herein by reference.

D¹-dehydration

Δ1-Dehydrogenation by DDQ in the presence of a silylated agent bistrimethylsilyltrihaloacetamide, hexamethyldisilane or bistrimethylsilylurea is described in U.S.P. 5,116,983, the disclosure of which is incorporated herein by reference.

15-alkyl compounds

The compounds of the invention in which 15-alkyl substitution is present may be prepared in a manner analogous to that described in PCT Application No. PCT/US94/02697 (WO 94/20114), the disclosure of which is incorporated herein by reference.

BIOLOGICAL METHODS & RESULTS

The following abbreviations are used below:

NADPH = hydrogenated nicotinamide adenine dinucleotide phosphate

DMSO = dimethyl sulfoxide

EDTA = ethylenediaminetetraacetic acid

In vitro C17,20-lyase assays: Compounds were tested for inhibition of cynomolgus monkey C17,20-lyase in vitro using microsomal preparations of the enzyme from testicular tissue. Testes were removed from anesthetized animals and snap frozen in liquid nitrogen. Microsomes were isolated as described in Schatzman et al., Anal. Biochem. 175, 219-226 (1988). The compound to be tested was dissolved in dimethyl sulfoxide and diluted in 0.05 M potassium phosphate buffer, pH 7.4, to obtain the desired concentrations of test compound in an amount that contributed 0.1% v/v DMSO to the total assay volume. Samples contained 0.05 M potassium phosphate buffer, pH 7.4, an NADPH-regenerating system (1 mM NADPH, 5 mM glucose-6-phosphate, 1 IU/ml glucose-6-phosphate dehydrogenase), test compound, substrate, and microsomal protein in a total volume of 0.2 ml. Control samples contained all components, including dimethyl sulfoxide, but no test compound. All assays were performed in duplicate. The test compound was incubated with 20 to 62 μg/ml microsomal protein, buffer, and the NADPH-regenerating system described above at 34°C for 0 or 40 minutes. Aliquots of 180 μl were then removed and assayed for enzyme activity by addition to 7-[³H]-17α-hydroxypregnenolone (11.2 mCi/mmol; 0.2 μCi per sample) plus unlabeled 17α-hydroxypregnenolone dissolved in DMSO, contributing 2.5% v/v to the final sample mixture, and phosphate buffer to give a total substrate concentration of 0.05 μM (=Km) per sample, followed by incubation at 34°C for 6 min. Each assay was terminated by addition of 5 mL chloroform:methanol (2:1). Carrier steroids (17α-hydroxypregnenolone, dehydroepiandrosterone, and androst-5-ene-3β,17β-diol), representing substrates and products, and 0.8 ml of distilled, deionized water were also added at this time. The steroids were extracted by the method of Moore and Wilson (Methods in Enzymol., eds. B. W. O'Malley and J. G. Hardman, 36, 1975, pp. 466-474). The organic phase containing the steroids was evaporated using nitrogen gas, the residue dissolved in 18% tetrahydrofuran (v/v) in hexane, and the steroids were separated by HPLC on a Si60 (5 µm) column (250 x 4 mm) using a gradient of 18-22% tetrahydrofuran (v/v) in hexane. Radioactivity in the steroid peaks was measured using a Radiomatic® detector model HS or model A515 Flo-One® detector.

Enzyme activity for each sample was calculated from the percent conversion of substrate to products and the results were expressed as percent inhibition of the control. The following results were obtained, where the values given are the mean of duplicate determinations: Table 1: In Vitro C17,20 Lyase Inhibition

LEGEND:

MDL 103,129 = 17β-cyclopropyloxy-4-aza-5α-androst-1-en-3-one

MDL 103,432 = 17β-cyclopropyloxy-4-aza-5α-androstan-3-one

MDL 103,496 = 17β-cyclopropyloxy-4-aza-androst-5-en-3-one

MDL 104,313 = 17β-Cyclopropyloxy-4-methyl-4-aza-androst-5-en-3-one

MDL 105,831 = 17β-cyclopropylamino-4-aza-androst-5-en-3-one

In vitro Soc reductase studies: The activity of the present compounds as inhibitors of steroid 5α-reductase was determined using microsomal preparations of the 5α-reductase enzyme from laboratory animal prostate tissue. Specifically, the microsomes were isolated from cynomolgus monkey prostate tissue. The protein concentration of the microsomal preparations was determined before use of the samples. Individual samples of cynomolgus monkey prostate 5α-reductase activity contained 0.1 M potassium phosphate-sodium citrate buffer, pH 5.6, 1.0% (w/v) bovine serum albumin, 1.0 ml sodium EDTA, 4 μg microsomal protein, 1.0 mM NADPH, 5.0 mlvi glucose-6-phosphate, 1 IU/ml glucose-6-phosphate dehydrogenase, [1,2-³H]-testosterone (0.15 μCi) plus unlabeled testosterone to give 0.015 μM (Km = 0.015 μM to 0.091 μM in duplicate determinations), and test compound dissolved in DMSO then dissolved in 0.1 M potassium phosphate-sodium citrate buffer, pH 5.6, to give a final sample concentration of 0.1% (v/v) DMSO. The same buffer and DMSO without test compound were used in control samples. Background radioactivity was determined from samples containing all components except the enzyme. Assays were performed in duplicate. Microsomes, 0.1 M potassium phosphate-sodium citrate buffer, pH 5.6, and test compound were preincubated at 37ºC. Aliquots of 180 µl were removed after 0 or 40 minutes of preincubation and added to 20 ml of testosterone substrate suspended in 0.1 M potassium phosphate-sodium citrate buffer, pH 5.6, containing 10% (v/v) DMSO. Remaining enzyme activity was assayed for 10 minutes at 37ºC in a Dubnoff shaking incubator.

Reactions were terminated by addition of 5 ml of CHCl3:methanol (2:1) and 0.9 ml of water. Carrier steroids were added as 2.5 µg each of testosterone, dihydrotestosterone and 3,17-androstanediol. Steroid metabolites were then extracted according to the method of Moore and Wilson (Methods in Enzymol., BW. O'Malley and JG Hardman, eds., 36, 1975, pp. 466-474), the organic phase containing the steroids was evaporated using nitrogen gas, the residue was dissolved in 3% (v/v) isopropanol in hexane. The steroids were then separated by normal phase HPLC on a LiCrosorb® DIOL derivatized silica gel column (10 µm; 4 x 250 mm) with a gradient of 3% to 7.5% isopropanol in hexane followed by isocratic conditions with 75% (v/v) isopropanol in hexane. Radioactivity in the steroid peaks was measured using a Packard Radiomatic Model LS Flo-One® detector. When the compounds were assayed using the above procedures with cynomolgus monkey 5α-reductase, the following results were obtained: Table 2: Results of in vitro 5α-reductase inhibition

*NOTE: Values are the mean of duplicate determinations See Table 1 for chemical names

Ex vivo inhibition of C17,20-lyase: MDL 103,432 was tested for ex vivo inhibition of testicular lyase of rats and nude mice. Male Copenhagen rats and male athymic nude mice obtained from Harlan Laboratories, Indianapolis, IN, were divided into groups of 5 to 6 based on weight. The average weight was 100-140 g each for rats and 18-35 g each for mice. Animals were fasted overnight prior to oral dosing. The test compound was prepared by micronization in a lecithin vehicle using a Teflon® pestle-type glass homogenizer. The compound was volumeted using lecithin such that 0.5 mL was administered per 100 g animal. Rats and nude mice were given vehicle alone (controls) or vehicle plus test compounds per os. Rats were also given the compound in lecithin or lecithin alone subcutaneously. Each group consisted of 5-6 animals. At a predetermined time after dosing, animals were anesthetized with CO2 gas, sacrificed by neck dislocation, testes removed, capsules removed, and tissue weighed. Two volumes (wt/vol) of 0.05 M potassium phosphate buffer, pH 7.2, were added to rat testis tissue on ice, and 11 volumes (wt/vol) of the same buffer were added to mouse testes. Tissue was then homogenized using 20 strokes with a Dounce homogenizer fitted with a tight-fitting pestle. The homogenized tissue was centrifuged for 15 minutes at 800 x g, then at 10,000 x g. The supernatant was decanted, saved and stored chilled on ice. Samples for lyase activity contained the same buffer and NADPH regenerating system described above and also contained 120 μl of the 10,000 x g supernatant diluted 3-fold. resulting in a total 5-fold dilution in the final sample volume. Substrate, 17α-hydroxyprogesterone plus 1,3-[³H]-17α-hydroxyprogesterone (40-57 mCi/mmol; 0.18 uCi per sample), giving a final concentration of 0.1 uM (=Km), was added to the remaining sample components after equilibration of the latter for 5 minutes at 20ºC. The total assay volume was 200 µl. Activity was assayed for 20 seconds at 20ºC.

Nude mouse testicular lyase was assayed by the same procedure as described for the rat enzyme above, except that the 10,000 x g supernatant was diluted 12-fold in phosphate buffer and 60 μl of this was used in the assay, resulting in a total 40-fold dilution of the supernatant. The substrate concentration was 0.04 μM (Km = 0.03 μM) and samples were incubated at 15°C for 30 seconds.

The assays were terminated, extracted and analyzed as described above, except that the carrier steroids were 17α-hydroxyprogesterone, androst-4-enedione and testosterone. The organic phase containing the steroids was evaporated using nitrogen gas, the residue dissolved in 18% tetrahydrofuran (v/v) in hexane, and the steroid substrate, 17α-hydroxyprogesterone and the products (AED, TEST) were separated by HPLC on a Si60 (5 µm) column (250 x 4 mm) using 20% (v/v) tetrahydrofuran (THF) in hexane for 20 minutes, then increasing to 60% THF (v/v) for 11 minutes. The activity of the test compound was expressed as percent inhibition relative to the control and was the mean value of the treated group of animals.

Using the method described above, MDL 103,432 inhibited the testicular C17,20-lyase activity of nude mice by 88% at 30 mg/kg and 96% at 100 mg/kg 4 hours after oral dosing. The testicular C17,20-lyase activity of rats was inhibited by MDL 103,432 as shown below: Table 3: Results of Ex Vivo C17,20-lyase Inhibition

In vivo data: Dunning H tumor

As described in J. T. Isaacs & D. S. Coffey, Cancer Res. 41: 5070-5075 (1981); W. J. Ellis & J. T. Isaacs, Cancer Res. 45: 6041-6050 (1985); T. W. Redding & A. V. Schally, The Prostate 6: 219-232 (1985) and P. E. Juniewicz et al. The Prostate 18: 105-115 (1991), male Copenhagen rats were obtained from HARLAN-SPRAGUE-DAWLEY Inc. (Indianapolis, IN) and were housed individually in suspended wire cages and fed laboratory rodent chow (Purina 5001 pellets, Purina Mills, St. Louis, MO) and deionized water ad libitum. Rats were anesthetized using sodium pentobarbital and hair from the posterior dorsal region was clipped. Tumors from donor Copenhagen rats were cut into 10 mm3 fragments and implanted subcutaneously (one site per rat) into the prepared dorsal region.

Animals were selected (105 days after implantation) for the treatment phase based on tumor size. Ten animals were anesthetized with sodium pentobarbital and bilaterally castrated. The remaining animals were assigned to treatment groups (ten per group) based on mean group tumor size. Animals were housed separately throughout the study. Tested compounds were prepared in solution or suspension in a lecithin vehicle (L-α-phosphatidylcholine type XV-E) containing methylparaben and propylparaben. All treatments were administered by oral gavage (per os) at 2 cc/kg each day of the study. Tumor size and rat body weights were recorded every seven days for 35 days. Twenty-four hours after the last treatment, animals were euthanized with CO2 and the tumors, prostate, seminal vesicles, and testes were removed and weighed. In Table 4, the average tumor growth is determined from the corrected group means over a 35-day period after treatment was initiated. The correction was determined by first omitting from each data set those animals that showed grossly disproportionate growth relative to the other animals in the treatment group. Since these tumors were indeed Raffen sarcomas and the phenomenon was observed in all compound-treated animals, such disproportionate growth is believed to be a limitation endemic to this model. The animals omitted from the calculation of corrected means were animals 8, 9, 10 in Fig. 6; animals 18, 19, and 20 in Fig. 7; in Fig. 8 animals 27, 28, 29 and 30 and in Fig. 9 animals 37, 39 and 40. The castrated controls in Fig. 10 present a different problem. Here it is believed that the different tumor volumes observed throughout the group are due to different sizes at the start of the study. Since the mean variance here is clearly due to other factors than can be attributed to the effect of castration itself, animals 47, 48, 49 and 50 were deleted from the corrected mean. Of these, animal 47 died during the treatment period.

Table 4 shows the average daily growth rate measured from the corrected mean tumor size on day 35 and day 0 comparing MDL 105831 and flutamide (MDL 15910), a known androgen receptor antagonist. MDL 105831 is shown to have similar tumor suppression properties to flutamide, which are additive when combination therapy is used. Table 4: Tumor growth in Dunning H rats (mm³)

LEGEND:

MDL 105831 = 17β-Cyclopropylamino-4-azaandrost-5-en-3-one (15 mg/kg/day p.o.)

MDL 15910 = N-(3-trifluoro-methyl-4-nitrophenyl)isobutyramide (50 mg/kg/day p.o.)

PC-82 tumors in nude mice:

Male nude mice (Hsd: athymic, Nude-nu) were obtained from Harlan Sprague Dawley as described in GJ von Steenbrugge et al., J. Urol. 131: 812-817 (1984), GJ von Steenbrugge et al., The Prostate 11: 195-210 (1987), and TW Redding et al., Cancer Research 52, 2538-2544 (1992). Mice were housed in sterilized microisolators and fed autoclaved ABLE® rodent chow (Purina Mills Inc., St. Louis, MO) and deionized water ad libitum. Tumor-donating mice were first anesthetized using sodium pentobarbital and then sacrificed by neck dislocation. The tumor was then removed and placed in a Petri dish containing ice-cold Hanks balanced salt solution. The tumors were cut into 2-3 mm cubes for implantation. The recipient animals were first anesthetized with 50 mg/kg pentobarbital, then tumor fragments (one per mouse) using a trocar in the dorsal region. Animals were separated into two control groups, one with vehicle alone and the other castrated, and treatment groups, where n is the number of animals in each group.

Animals were selected for treatment groups based on tumor size. Each test compound was prepared as a solution or suspension in a lecithin vehicle (L-α-phosphatidylcholine type XV-E) containing methylparaben and propylparaben at a dose volume of 10 cc/kg. Animals were treated by oral gavage (per os) seven days per week for 42 days. Twenty-four hours after the last treatment, animals were euthanized with CO2 and tumors were removed and weighed. During the study period, mice were weighed and palpated weekly for tumors. In Table 5, average tumor growth is determined from corrected group means over a 28-day period after treatment was initiated. Correction was determined by first removing from each data set those animals that showed grossly disproportionate growth relative to the other animals in the treatment group. Such tumor growth is believed to result from conversion of the tumor to a non-androgen-dependent sarcoma and most frequently resulted in euthanasia of the experimental animal before the end of the treatment period. The following animal data were deleted before calculation of the corrected mean data in Table 5: Figure 1: Animals 4 and 44; Figure 2: Animals 10, 38 and 42; Figure 3: Animal 39; Figure 4: Animals 8, 9 and 49; Figure 5: Animals 37 and 40 and 41.

Table 5 illustrates the average tumor growth in animals treated with 4-aza-17β-(cyclopropyloxy)-5α-androstan-3-one (MDL 103432; 50 mg/kg B. LD.), 4-aza-17β-(cyclopropylamino)-androst-5(6)-en-3-one (MDL 105831; 50 mg/kg B. LD.), and flutamide (MDL 15910; 15 mg/kg B. LD.), a known androgen receptor antagonist. The average growth rate of each compound tested relative to vehicle and castrated controls is consistent with in vivo inhibition of androgens. Table 5: Tumor growth of human PC-82 tumor in male nude mice (mm³)

EXAMPLES

The following examples are given to better illustrate the syntheses of the particular compounds of the invention and should not be construed in any way as a limitation of the invention.

DEFINITIONS

Unless otherwise noted in the following examples, "room temperature" means 18ºC-23ºC, any mention of "overnight" means 14-18 hours, and dissolved reagents are in aqueous solutions. The following formula abbreviations have also been used:

Saline solution = saturated aqueous solution of sodium chloride (NaCl)

THF = Tetrahydrofuran

EtOAc = ethyl acetate

MgSO₄ = Magnesium sulfate

HOAc = acetic acid

Na&sub2;SO&sub3; = sodium sulfite

NaHCO3 = sodium bicarbonate

CH2Cl₂ = methylene chloride

Ether = diethyl ether (CH3 CH2 )2 O

NH₄Cl = ammonium chloride

EXAMPLE 1 17β-Cyclopropyloxy-4-azaandrost-5(6)-en-3-one EXAMPLE 1A 17β-Hydroxy-5-oxo-4-nor-3,5-seco-androstane-3-carboxylic acid

By a procedure analogous to that described by L. Milewich & L. Axerrod, Organic Synthesis, Collect. Vol. 6, 1988, 690 - 91, testosterone (UPJOHN, 9.16297 g, 31.772 mmol) is dissolved in tert-butyl alcohol (300 mL) in a 1000 mL three-necked round-bottom flask. Potassium carbonate (K2CO3) in water (75 mL) is added and stirred until completely dissolved. The reaction flask is equipped with a 500 mL dropping funnel charged with sodium metaperiodate (NaJO4, ALDRICH, 40.9249 g, 191.34 mmol) in water (350 mL). The reaction flask is also equipped with a separate 125 mL dropping funnel charged with a solution of potassium permanganate (FLUKA, KMnO4, 0.80424 g, 5.089 mmol) in water (50 mL).

About 50 mL of the metaperiodate solution and about 5 mL of the permanganate solution are added to the reaction mixture in a single batch. The remainder of each solution is added dropwise over 30 minutes. After the additions are complete, the reaction mixture is stirred for an additional 90 minutes. The reaction is then quenched by slowly adding potassium bisullite (K2S2O5, BAKER, 23.70107 g, 106.603 mmol) and stirring for 5 hours.

The reaction mixture is then filtered through Celite® filter aid and stored overnight at room temperature.

The filtrate is concentrated under reduced pressure (45 mm Hg, 70°C) to about 250 mL and transferred to a 500 mL separatory funnel. The concentrate is acidified with 10% sulfuric acid (H2SO4, 26 mL) and extracted with ether (3 x 200 mL). The combined ethereal extracts are washed with 100 mL of diethyl ether, then poured into 10% sulfuric acid (300 mL) to precipitate the product.

The precipitate is extracted with methylene chloride (CH2Cl2) (4 x 200 mL), the organic layers are combined and washed with brine (100 mL), dried over MgSO4, filtered and evaporated to dryness to give a white solid. The material is recrystallized from acetone (ca. 50 mL) overnight, collected by filtration, washed in hexane (30 mL) and dried under reduced pressure (0.3 mm Hg, room temperature) for 4.5 hours to give 17β-hydroxy-5-oxo-4-nor-3,5-seco-androstane-3-carboxylic acid (6.1247 g). The compound has the following structure:

Alternatively, the 4-nor-3,5-seco acid can be prepared as follows: Testosterone (9.1863 g) is dissolved in CH2Cl2 (70 mL) and diluted with methanol (100 mL). The solution is quenched to -78°C under nitrogen. Ozone is then bubbled through the quenched solution for 25 minutes, after which the solution turns green. The atmosphere of the reaction vessel is purged with nitrogen, the reaction mixture is warmed to room temperature and the solvent is evaporated. The residue is dissolved in ether (200 mL), extracted three times with 10% sodium hydroxide solution (NaOH, 50 mL), washed and the organic phases are combined and washed again in ether (50 mL) and acidified with 10% sulfuric acid (H2SO4, 200 mL). The acidified solution is extracted into CH2Cl2 (4 x 50 mL) and the combined organic layers are washed with brine, dried over MgSO4, filtered and evaporated to give the crude product as a white foam.

The crude product is taken up in hot acetone (100 mL, 50ºC) and concentrated (~40 mL). The colorless crystals that form after cooling are collected and analyzed for purity to give 17β-hydroxy-5-oxo-4-nor-3,5-seco-androstane-3-carboxylic acid.

EXAMPLE 1B 17β-Acetoxy-4-azaandrost-5(6)-en-3-one

In a manner analogous to that disclosed in M. Kobyashi & H. Mitsuhashi, Chem. Pharm. Bull., 1973, 21 (5), 1069-1075, the seco acid prepared above (4.0249 g, 13.051 mmol) and ammonium acetate (NH4OAc, EM SCIENCE, 10.26 g, 133.1 mmol) are dissolved in HOAc (130 mL) and heated to reflux temperature under nitrogen. After refluxing for 5 days, the reaction mixture is cooled to room temperature, then poured into water (800 mL). The resulting precipitate is collected by filtration and dried overnight at reduced pressure and room temperature (0.3 mm Hg).

The precipitate is recrystallized from ethanol by adding it to boiling ethanol (350 mL), concentrating it (~180 mL) and drying it (reduced pressure, room temperature) to give 17β-acetoxy-4-azaandrost-5(6)-en-3-one, which corresponds to the chemical formula:

EXAMPLE 1C 17β-Hydroxy-4-azaandrost-5(6)-en-3-one

The 17β-ethyl carboxylate (0.9259 g, 2.793 mmol) prepared in Example 1B is dissolved in 60 mL of warm ethanol/tetrahydrofuran (1:1), 6 M sodium hydroxide solution (NaOH, 10.0 mL, 60 mmol) is added, and the reaction mixture is stirred at room temperature for 2 1/2 hours. The reaction mixture is diluted with brine (100 mL) and extracted with EtOAc (3 x 50 mL). The combined organic layers are washed with brine (50 mL) and ammonium chloride (NH4Cl), dried over magnesium sulfate, filtered, and evaporated to give a crude reaction product. The crude product is recrystallized from ethanol to give colorless crystals (0.3132 g, 1.0822 mmol) of 17β-hydroxy-4-azaandrost-5(6)-en-3-one. The mother liquor is concentrated and the above recrystallization procedure is repeated to give a second crop of crystals (0.3945 g, 1.363 mmol).

EXAMPLE 1D 17β-Vinyloxy-4-azaandrost-5(6)-en-3-one

The 17β-alcohol prepared in Example 1C (2.20 g, 7.77 mmol) is made into a slurry with chloroform (CHCl3, 30 mL) and ethyl vinyl ether (CH2CHOC2H5, 40 mL). Mercury(11) acetate (ALDRICH, Hg(OOCCH3)2, 2.4869 g, 7.804 mmol) is added to the reaction vessel, which is then purged with nitrogen, and heated to reflux under a nitrogen atmosphere. After refluxing for 14 1/2 hours, when the reaction mixture is dark brown and homogeneous, the reaction is quenched with acetic acid (HOAc, 0.20 mL, 0.21 g, 3.49 mmol) and stirred at room temperature for an additional 2 1/2 hours. The reaction mixture is poured into 5% sodium hydroxide solution (50 ml) and hexane (150 ml), the phases are separated and the organic phase is washed with brine (2 x 50 mL), dried over potassium carbonate (K2CO3), filtered and evaporated to give 17β-vinyloxy-4-azaandrost-5(6)-en-3-one (5.033 g). The compound has the following structure:

EXAMPLE 1E 17β-Cyclopropyloxy-4-azaandrost-5(6)-en-3-one

In a manner analogous to the procedure described in Charette et al., Tet. Let., 1994, 35(4), 513-16, the vinyl ether (7.77 mmol) is dissolved in chloroform (CHCl3) under nitrogen. Tert-butyl methyl ether (CH30C(CH3)3, 20 mL) is added, initiating the formation of an off-white precipitate. The mixture is quenched to 0°C under nitrogen. Diethylzinc ((CH3CH2)2Zn), ALDRICH, 24.0 mL, 26.4 mmol, 1.1 M in toluene) is added to the vinyl ether slurry, partially dissolving the precipitate. The slurry is stirred at 0°C for 10 minutes and methylene iodide (CH2I2, ALDRICH, 2.20 mL, 7.32 g, 27.3 mmol) is added in small portions over 15 minutes and the slurry is continuously stirred at 0°C under nitrogen. Another portion of chloroform (CHCl3, 40 mL) is added and stirring is continued at 0° for a total reaction time of 6 hours. The reaction mixture is poured into 120 mL of a saturated NH4Cl solution and extracted with EtOAc (200 mL). The organic layer is extracted with brine (2 x 100 mL), dried over MgSO4 dried, filtered and evaporated to give the crude product which, when purified by flash chromatography on silica (SiO2, RF 0.29) with ethyl acetate/methylene chloride/hexane eluent (25% EtOAc/25% CH2Cl2/50% hexane), gave 17β-cyclopropyloxy- 4-azaandrost-5(6)-en-3-one (1.3002 mmol, 17% yield).

1 H-NMR (300 MHz, CDCl3 ) ? 8.78 (br s, 1H), 4.93 (dd, J = 4.9; 2.0 Hz, 1H), 3.45 (t, J = 8.4 Hz, 1H), 3.26- 3.35 (m, 1H), 2.42-2.51 (m, 2H), 1.85-2.21 (m, 4H), 1.37-1.69 (m, 7H), 1, 10 (s, partially hidden, 3H), 0.94-1.36 (m, 4H), 0.79 (s, 3H), 0.40-0.60 (m, 4H);

13 C-NMR (75 MHz, CDCl3 ) ? 169.9; 139.9; 103.3; 88.9; 52.3; 51.2; 48.0; 42.5; 37.1; 34.0; 31.5; 31.4; 29.2; 28.3; 27.8; 23.2; 20.5; 18.6; 11.6; 6.1; 5.8;

IR (KBr) 3435 (br), 2967 (m), 1669 (s) cm⁻¹;

MS (electron impact) m/e calcd for C₂₁₁H₃₂NO₂: 330.243305; found: 330.242958; 329 (molecular ion), 314, 288, 272 (base), 244, 230, 176, 162, 138, 135, 108.

The compound has the following structure:

EXAMPLE 2 17β-Cyclopropyloxy-4-aza-5α-androstan-3-one EXAMPLE 2A 17β-Acetoxy-4-aza-5α-androstan-3-one

17β-Acetoxy-4-aza-5α-androst-5(6)-en-3-one (10.0404 g, 30.2914 mmol) and palladium/carbon catalyst (ENGELHARD, 1.2642 g, 5% Pd on carbon) are placed in a 500 mL Parr bottle. The reaction vessel is purged with nitrogen and ethanol (250 mL) is added. The reaction vessel is then charged with hydrogen to 60 psi and heated to 60ºC with mechanical stirring. At the elevated temperature, the pressure of the reaction vessel increases, but then decreases after reaction with the steroid. The hydrogen pressure is maintained around 60 psi by periodic gas additions through a ballast vessel until the hydrogen pressure becomes constant (about 90-100 hours). After completion of the reaction, the reaction mixture is cooled to room temperature, washed with acetic acid (70 mL) through Celite®, filtered, and concentrated to dryness to give the crude product. The crude product is recrystallized by dissolving in boiling solvent (400 mL) and concentrating from ethanol (150 mL). The colorless crystals are collected and dried overnight under reduced pressure (0.4 mm Hg) at room temperature and analyzed for purity to give 17β-acetoxy-4-aza-5α-androstan-3-one. Yield: 25.08 mmol, 83%. The compound has the following structure:

EXAMPLE 2B 17β-Hydroxy-4-aza-5α-androstan-3-one

The 17β-acetoxy-4-aza-5α-androstan-3-one prepared in Example 2A (2.6533 g, 7.9559 mmol) is dissolved in warm ethanol (100 mL). Concentrated sodium hydroxide solution (6 M, 50 mL) is added and the reaction mixture is stirred at room temperature. After 90 minutes, the reaction mixture is worked up by first diluting the reaction mixture with brine (200 mL) and extracting with EtOAc (3 x 100 mL). The combined organic layers are washed with brine (100 mL), dried over MgSO4, filtered and evaporated to give 17β-hydroxy-4-aza-5α-androstan-3-one.

EXAMPLE 2C 17β-Vinyloxy-4-aza-5α-androstan-3-one

In a manner analogous to that described in Ireland et al., Org. Synth., Coll. Vol. VI, 1988, 298-301, a slurry is prepared from 4-aza-17β-vinyloxyandrostan-3-one (0.5297 g, 1.817 mmol) with CH2Cl2 (15 mL) and ethyl vinyl ether (CH3CH2OCHCH2, ALDRICH, 11.31 g, 156.8 mmol). Mercury(II) acetate (Hg(OOCCH3)2, ALDRICH, 0.6030 g, 1.8922 mmol) is added and the slurry is heated to reflux under nitrogen for 39 hours. Glacial acetic acid (HOOCCH3, EM, 0.050 mL, 0.053 g, 0.873 mmol) is added and the reaction mixture is cooled to room temperature while stirring overnight. The reaction mixture is worked up by pouring it into 5% aqueous sodium hydroxide (15 mL) and hexane (40 mL). The layers are separated and the organic layers are washed with brine (2 x 24 mL). The combined organic layers are washed, filtered and evaporated to give 17β-vinyloxy-4-aza-5α-androstan-3-one (1.0581 g), which is used immediately in the following step without further characterization.

EXAMPLE 2D 17β-Cyclopropyloxy-4-aza-5α-androstan-3-one

The 4-aza-17β-vinyloxy-5α-androstan-3-one (1.817 mmol) prepared in Example 2C is dissolved in a cosolvent system of CH2Cl2 (12 mL) and methyl tert-butyl ether (12 mL) and quenched to 0°C under nitrogen. Diethylzinc ((CH3CH2)2Zn) is added, followed by methylene iodide (CH2I2, ALDRICH, 1.80 mL, 5.96 g, 22.3 mmol) added in small portions over 2 minutes. The mixture is stirred at 0°C under nitrogen for 6 1/4 hours. The reaction was quenched with saturated aqueous NH4Cl (20 mL) and extracted with EtOAc (100 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over MgSO4, filtered and the solvents were evaporated to give a crude product of the title compound. The crude product was purified by flash chromatography (SiO2, 50% EtOAc/50% CH2Cl2) and the product-containing fraction was collected to give pure 17β-cyclopropyloxy-4-aza-5α-androstan-3-one (0.4223 g, 1.2739 mmol, yield: 70%). mp 237.5-238.5 °C (from acetone).

RF 0.16 (50:50 CH2 Cl2 :EtOAc);

1 H-NMR (300 MHz, CDCl3 ) ? 6.70 (br s, 1H), 3.43 (t, J = 8.4 Hz, 1H), 3.24-3.33 (m, 1H), 3.04 (dd, J = 12.1 ; 3.6 Hz, 1H), 2.35-2.43 (m, 2H), 1.92-2.05 (m, 1H), 1.82-1.90 (m, 2H), 1, 71 (dd, J = 12.9; 2.9 Hz, 1H), 1.10-1.59 (m, 11H), 0.90 (s, 3H), 0.85-1.03 (m, partially hidden, 2H), 0.76 (s, 3H), 0.41-0.59 (m, 4H);

13 C-NMR (75 MHz, DMSO-d6 ) ? 170.1; 88.3; 59.7; 51.9; 50.7; 50.0; 42.3; 37.0; 35.0; 34.4; 32.9; 28.7; 28.4; 27.5; 26.2; 22.8; 20.3; 11.7; 11.1; 5.8; 5, 6;

IR (KBr) 3434 (br), 3194 (m), 1672 (s) cm⁻¹;

MS (electron impact) m/e calcd for C₂₁₁H₃₄NO₂: 332.258955; found 332.257883; 331 (molecular ion), 315, 288, 274 (base), 191, 163, 124, 112.

The product has the following formula:

EXAMPLE 3 17β-Cyclopropyloxy-4-methyl-4-azaandrost-5(6)-en-3-one EXAMPLE 3A 17β-Acetoxy-4-methyl-4-azaandrost-5(6)-en-3-one

From 17β-hydroxy-5-oxo-4-nor-3,5-seco-androstane-3-carboxylic acid prepared in Example 1A or otherwise obtained (3.1856 g, 10.329 mmol), a slurry is prepared with methylamine hydrochloride (CH3NH2·HCl, ALDRICH, 7.4847 g, 110.9 mmol) and HOAc (80 mL) and heated to reflux under nitrogen. After six days, the reaction mixture is cooled to room temperature and the reaction mixture is concentrated (30 mm Hg, 55°C) to give a thick slurry. The slurry is taken up in EtOAc (100 mL) and the organics are washed with water (2 x 50 mL), with aqueous NaHCO3 solution (50 mL) and with brine (50 mL). The product is dried over MgSO4, filtered and the solvent is evaporated to give the crude product which is then purified by flash chromatography (SiO2, 25% EtOAc/25% CH2Cl2/50% hexane) to give 17β-acetoxy-4-methyl-4-azaandrost-5(6)-en-3-one (3.3373 g, 9.763 mmol). Yield: 95%. The compound has the following structure:

EXAMPLE 3B 17β-Hydroxy-4-methyl-4-azaandrost-5(6)-en-3-one

17β-Acetoxy-4-methyl-4-azaandrost-5(6)-en-3-one (3.373 g, 9.763 mmol) was dissolved in ethanol (50 mL). Concentrated sodium hydroxide solution (6 M, 25.0 mL) was added and the reaction mixture was stirred at room temperature for 4 1/2 hours. The reaction mixture was partitioned between brine (10.0 mL) and EtOAc (100 mL). The layers were separated and the organic layer was washed with brine (2 x 100 mL) and saturated NH4Cl solution (100 mL), dried over MgSO4, filtered and evaporated. to give a crude reaction product as a yellow solid. This is purified by dissolving in methanol and evaporating until the formation of large crystals is observed. The crystals are washed in acetone and dried overnight under reduced pressure (0.3 mm Hg) to give 17β-hydroxy-4-methyl-4-azaandrost-5(6)-en-3-one of very good purity (1.8119 g, 5.9712 mmol). Yield: 61%. M.P. 188ºC-192ºC.

EXAMPLE 3C 4-Methyl-17β-vinyloxy-4-azaandrost-5(6)-en-3-one

17β-Hydroxy-4-methyl-4-azaandrost-5(6)-en-3-one (0.9278 g, 3.0576 mmol) was dissolved in CH2Cl2 to which ethyl vinyl ether (CH3CH2OCHCH2, ALDRICH, 20.0 mL, 15.08 g, 209.1 mmol) was added while stirring. Mercury(II) acetate (Hg(OOCCH3)2) was added and the reaction mixture was heated to reflux under nitrogen. After 24 1/2 hours, the reaction mixture was cooled to room temperature and HOAc (EM, 1.00 mL, 1.06 g, 17.46 mmol) was added. After 15 1/2 hours, the reaction mixture is poured into a mixture of hexane (100 mL) and 10% sodium hydroxide solution. The container is mixed well, then the phases are separated and the organics are washed with brine (2 x 50 mL), dried over MgSO4, filtered and the solvent is evaporated to give 4-methyl-17β-vinyloxy-4-azaandrost-5(6)-en-3-one (1.3829 g) which corresponds to the structure:

EXAMPLE 3D 17β-Cyclopropyloxy-4-methyl-4-azaandrost-5(6)-en-3-one

The vinyl ether prepared in Example 3C (4-methyl-17β-vinyloxy-4-azaandrost-5(6)-en-3-one, 3.05 mmol) is dissolved in a mixture of CH2Cl2 (20 mL) and t-butyl methyl ether (20 mL) and quenched to 0°C under nitrogen. Diethylzinc ((CH3CH2)2Zn, AL-DRICH, 20.0 mL, 1.1 M in toluene, 22.0 mmol) is added, followed by addition of methylene iodide (CH2I2, ALDRICH, 1.80 mL, 5.96 g, 22.3 mmol) in small portions. After 6 1/2 hours, the reaction mixture was worked up by first quenching with saturated NH4Cl solution (50 mL) and extracting with EtOAc (100 mL). The organic layers were washed with brine (2 x 50 mL), dried over MgSO4, filtered and the solvent was evaporated to give the crude reaction product. The crude product is purified by flash chromatography (SiO2, 25% EtOAc/25% CH2Cl2/50% hexane) to give 4-methyl-17β-cyclopropyloxy-4-azaandrost-5(6)-en-3-one (0.4095 g, 1.1921 mmol) which, when further analyzed, shows the presence of some minor impurities.

Rf0.48 (50:25:25-hexane:CH2 Cl2:EtOAc)

1 H-NMR (300 MHz, CDCl3 ) ? 5.04 (br d, J = 3.6 Hz, 1H), 3.46 (t, J = 8.4 Hz, 1H), 3.27- 3.35 (m, 1H), 3.12 ( s, 3H), 2.53 (br ABq, J = 3.6 Hz, 2H), 2.24 (dt, J = 16.8; 5.0 Hz, 1H), 1.87-2.09 ( m, 3H), 1.40-1.73 (m, 6H), 1.06 (s, 3H), 0.98-1.34 (m, partially hidden, 5H), 0.79 (s, 3H ), 0.42-0.61 (m, 4H);

13 C-NMR (75 MHz, CDCl3 ) ? 168.1; 144.0; 104.1; 88.7; 52.2; 51.0; 48.8; 42.3; 37.1; 35.2; 31.5; 30.9; 30.6, 30.1; 28.7; 27.7; 23.1; 20.4; 18.6, 11.5; 6.0; 5.7;

IR (KBr) 3437 (br), 1676 (s), 1647 (s) cm⁻¹;

MS (electron impact) m/e 343 (molecular ion), 328, 302, 286 (base), 270, 244, 190, 176, 152, 135, 124.

EXAMPLE 4 17β-Cyclopropyloxy-4-aza-5α-androst-1-en-3-one EXAMPLE 4A 17β-Acetoxy-4-aza-5α-androst-1-en-3-an

In a manner analogous to the procedure described in A. Bhattacharya et al., J. Am. Chem. Soc., 1988, 110, 3318-3319, 17β-acetoxy-4-aza-5α-androstan-3-one (0.7687 g, 2.3049 mmol) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ, ALDRICH, 0.5461 g, 2.4056 mmol) is dissolved in 1,4-dioxane (15.0 mL) under nitrogen. Bis(trimethylsilyl)trifluoroacetamide (BSTFA, ALDRICH, 2.60 mL, 2.52 g, 9.79 mmol) is added and the reaction mixture is stirred at room temperature for 60 min. The reaction mixture is heated to reflux for 18 h, cooled to room temperature and the solvent is allowed to evaporate. The residue is dissolved in EtOAc (50 mL), washed with 5% sodium hydroxide solution (30 mL) and brine (50 mL), dried over MgSO4, filtered and the solvent is evaporated to give the crude reaction product. The crude product is purified by flash chromatography (SiO2, 50% EtOAc/50% CH2Cl2) to give 17β-acetoxy-4-aza-5α-androst-1-en-3-one (0.6102 g, 1.8409 mmol) of very good purity. Yield: 80%. The compound has the following formula:

EXAMPLE 4B 17β-Hydroxy-4-aza-5α-androst-1-en-3-one

17β-Acetoxy-4-aza-5α-androst-1-en-3-one (0.500 g, 1.508 mmol) is dissolved in warm ethanol (20 mL). 6 M sodium hydroxide solution (10 mL) is added and the reaction mixture is stirred at room temperature for 16 hours. The reaction mixture is diluted with EtOAc (50 mL), extracted with brine (3 x 100 mL), and dried over MgSO4 to give 17β-hydroxy-4-aza-5α-androst-1-en-3-one (0.4570 g) and a minor impurity. The reaction product is carried to the next synthesis without further purification.

EXAMPLE 4C 17β-Vinyloxy-4-aza-5α-androst-1-en-3-one

In a manner analogous to that in Examples 1D, 2C and 3C and as described in RE Ireland et al., Org. Synth., Coll. Vol. VI, 1988, 298-301, 17β-hydroxy-4-aza-5α-androst-1-en-3-one (1.508 mmol), ethyl vinyl ether (CH₃CH₂OCHCH₂; ALDRICH, The reaction was reacted with acetic acid (EM, 0.346 g, 5.76 mmol) and mercury(II) acetate (Hg(OAc)2, ALDRICH, 0.5101 g, 1.6007 mmol) in CH2Cl2 (15 mL) for 66 hours. The reaction was quenched with acetic acid (EM, 0.346 g, 5.76 mmol) and stirred for 1 hour. The reaction mixture was diluted with 10% sodium hydroxide solution (25 mL), extracted with brine (2 x 25 mL), filtered and dried to give 17β-vinyloxy-4aza-5α-androst-1-en-3-one (0.6906 g) in quantitative yield, which was carried to the next step without further purification.

EXAMPLE 4D 17β-Cyclopropyloxy-4-aza-5α-androst-1-en-3-one

In a manner analogous to that in Examples 1E, 2D, and 3D, 17β-vinyloxy-4-aza-5α-androst-1-en-3-one (1.508 mmol), tert-butyl methyl ether (10 mL), diethylzinc (ALDRICH, 1.1 M in toluene, 10.00 mL, 11.00 mmol), and methylene iodide (CH2I2, ALDRICH, 0.900 mL, 2.99 g, 11.17 mmol) are reacted with the CH2I2 additions occurring in small portions over 5 minutes. After 2 1/2 hours, the reaction mixture was brought to room temperature and after 14 hours was worked up by quenching with saturated NH4Cl solution (50 mL), extracting with EtOAc (75 mL) and washing with brine (70 mL). This product was purified with 50% EtOAc/50% CH2Cl2 eluent to give 17β-cyclopropyloxy-4-aza-5α-androst-1-en-3-one (0.2246 g, 0.6816 mmol) in good purity. Yield: 45%.

RF 0.41 (50:50-CH2 Cl2 :EtOAc).

1 H-NMR (300 MHz, CDCl3 ) ? 6.79 (d, J = 10.0 Hz, 1H), 6.62 (br s, 1H), 5.80 (dd, J = 9.9; 2.2 Hz, 1H), 3.45 ( t, J = 8.4 Hz, 1H), 3.25-3.34 (m, 2H), 1.92-2.05 (m, 2H), 1.31-1.77 (m, 8H) , 1.13-1.30 (m, 3H), 0.97 (s, 3H), 0.90-1.07 (m, partially hidden, 2H), 0.77 (s, 3H), 0, 41-0.59 (m, 4H).

13 C-NMR (75 MHz, CDCl3 ) ? 166.8; 150.9; 122.8; 88.7; 59.5; 52.3; 50.5; 47.6; 42.8; 39.2; 37.2; 35.0; 28.9; 27.8; 25.6; 23.1; 20.8; 11.9; 11.8; 6.0; 5.8.

IR (KBr) 3425 (br), 3200 (m), 1682 (s) cm⁻¹;

MS (electron impact) m/e calcd Their C₂₁H₃₂NO₂: 330.243305; found 330.243780; 329 (molecular ion), 314, 286, 272 (base), 256, 228, 190, 163, 148, 122, 110.

The compound has the following structure:

EXAMPLE 5 17β-Cyclopropylamino-4-azaandrost-5(6)-en-3-one EXAMPLE 5A 5,17-Dioxo-4-nor-3,5-seco-androstane-3-carboxylic acid

Testosterone (8.7813 g, 30.45 mmol) is dissolved in a mixture of CH2Cl2 (50 mL) and methanol (10 mL) and the solution is quenched to -78°C under nitrogen while stirring. Ozone is bubbled through the reaction mixture while maintaining the temperature and stirring for 3 hours. Note that the reaction color turns blue after 30 minutes. The reaction mixture is slowly brought to room temperature while replacing the ozone with nitrogen. The solvent is evaporated and the residue is taken up in ether (200 mL) and extracted with 10% sodium hydroxide solution (3 x 50 mL), noting gas evolution. The alkaline layers are combined and washed again with ether (50 mL). The solution is acidified with 10% sulfuric acid (H2SO4, 200 mL) and extracted with CH2Cl2 (5 x 50 mL). The organic layers are combined and extracted with brine (50 mL), dried over MgSO4, filtered and the solvent is evaporated to give crystals of 5,17-dioxo-4-nor-3,5-seco-androstane-3-carboxylic acid (8.14 g).

EXAMPLE 5B 4-Azaandrost-5(6)-en-3,17-dione

The seco acid prepared in Example 5A (8.14 g, 26.56 mmol) is dissolved in HOAc (100 mL). Ammonium acetate (NH₄OOCCH₃, 15.6931 g, 203.6 mmol) is added and the reaction mixture is heated to reflux under nitrogen. After 69 hours, the reaction mixture is cooled to room temperature and poured into ice water (700 mL), whereupon the product separates as a tarry mass. The solution is washed with brine (200 mL) and extracted with EtOAc (3 x 200 mL). Combine the organic layers and wash with brine (2 x 200 mL), water (3 x 200 mL) and saturated NaHCO3 solution (2 x 100 mL). Dry the extract over MgSO4, filter and evaporate the solvent to give the crude title compound. Recrystallize the product from ethanol to give 4-azaandrost-5(6)-ene-3,17-dione (1.0859 g, 3.778 mmol) with traces of the C17 acetate. A second recrystallization of the mother liquor affords additional compound (0.5431 g). Overall yield: 21%. The desired product has the following structure:

EXAMPLE 5C 17β-Cyclopropylamino-4-azaandrost-5(6)-en-3-one

4-Azaandrost-5(6)-ene-3,17-dione (0.53 g, 1.844 mmol) was dissolved in chloroform (CHCl3, 10.0 mL) and cyclopropylamine ((CH2)2CHNH2, ALDRICH, 4.12 g, 72.15 mmol) was added and the reaction mixture was heated to reflux under nitrogen. After 15 h, the reaction mixture was cooled to room temperature and analysis showed stoichiometric conversion to the cyclopropylimine. Sodium borohydride (NaBH4, ALPHA, 0.3574 g, 0.4475 mmol) in ethanol (21 mL) was added and the reaction mixture was stirred at room temperature under nitrogen. After 21 hours, the reaction mixture was worked up by diluting with EtOAc (50 mL), washing with water (2 x 40 mL) and brine (40 mL), drying over MgSO4, filtering and evaporating the solvent to give the crude title compound. The crude product is further purified by flash chromatography (10.5", 5.25 g, SiO2; first eluting with 25% EtOAc/25% CH2Cl2/50% hexane, then with 15% i-PrOH/85% CH2Cl2) to afford 17β-cyclopropylamino-4-azaandrost-5(6)-en-3-one in excellent purity (0.4948 g, 1.5063 mmol).

Yield: 82%.

RF 0.42 (15:85-i-PrOH:CH2 Cl2 );

1 H-NMR (300 MHz, CDCl3 ) ? 9.22 (br s, 1H), 4.93-4.97 (m, 1H), 2.67 (t, J = 8.5 Hz, 1H), 2.42-2.51 (m, 2H ), 1.86-2.22 (m, 5H), 1.40-1.70 (m, 7H), 1.09 (s, 3H), 1.04-1.40 (m, partially hidden, 5H), 0.73 (s, 3H), 0.28-0.44 (m, 4H).

13 C-NMR (75 MHz, DMSO-d6 ) ? 167.7; 140.7; 101.0; 68.7; 52.8; 47.8; 42.1; 37.2; 33.5; 31.2; 31.1; 29.6; 29.0; 28.9; 28.2; 23.3; 20.2; 18.4; 11.6; 6.9; 6.3.

IR (KBr) 3429 (br), 3202 (m), 1669 (s) cm⁻¹.

MS (electron impact) m/e 328 (molecular ion), 313, 299 (base), 271, 256, 243, 228, 204, 162, 137, 108.

The compound has the following structure:

EXAMPLE 6 17β-Cyclopropylamino-4-aza-5α-androstan-3-one EXAMPLE 6A 5,17-Dioxo-4-nor-3,5-seco-androstane-3-carboxylic acid

Androst-4-ene-3,17-dione (10.000 g, 34.914 mmol) is dissolved in a mixture of CH2Cl2 (100 mL) and EtOAc (100 mL) and the solution is quenched to -78 °C under nitrogen. Ozone is bubbled through a coarse glass frit under the surface of the solution until the reaction mixture turns dark blue. The reaction mixture at -78 °C is then sparged with dry nitrogen until the blue color disappears. The solution is allowed to warm to ambient temperature and the solvent is removed under reduced pressure. The residue is taken up in ether (250 mL) and the product is extracted with 10% sodium hydroxide solution (3 x 25 mL). The combined basic extracts are washed with fresh ether (100 mL), then acidified with 10% sulfuric acid (H2SO4; 100 mL). The acidic aqueous solution is then extracted with CH2Cl2 (4 x 50 mL). The combined CH2Cl2 extracts are washed once with brine (50 mL), dried over MgSO4, filtered and the solvent is evaporated under reduced pressure to give the title compound.

EXAMPLE 6B 4-Azaandrost-5(6)-en-3,17-dione

5,17-dioxo-4-nor-3,5-seco-androstane-3-carboxylic acid prepared in Example 6A (9.000 g, 29.373 mmol) and ammonium acetate (NH4OAc; 22.58 g, 292.9 mmol) are slurried in HOAc (75 mL) and heated to reflux under an inert atmosphere. After 3 days, the reaction mixture is allowed to cool to room temperature and then poured into ice-cold water (700 mL). The resulting precipitate is collected by filtration, air-dried, and recrystallized from ethanol to give the title compound.

EXAMPLE 6C 17β-Hydroxy-4-aza-5α-androstan-3-one

4-Azaandrost-5(6)-ene-3,17-dione (7.000 g, 24.356 mmol) (see Example 6B) and 5% palladium on carbon catalyst (0.750 g, 0.352 mmol Pd) are placed in a 500 mL Parr bottle under an inert atmosphere. Acetic acid (100 mL) is added to the reaction vessel, which is then charged with hydrogen to 60 p.s.i. The hydrogenation reaction mixture is heated to 60 °C and agitated with a Parr shaker. After three days, the reaction mixture is allowed to cool to ambient temperature and filtered through Celite®. The filtrate is concentrated under reduced pressure to about 25 mL, then poured into ice-cold water (300 mL). The precipitate is then collected by filtration and recrystallized from ethanol to give the title compound.

EXAMPLE 6D 4-Aza-5α-androstane-3,17-dione

The 17β-hydroxy-4-aza-5α-androst-3-one (6.000 g, 20.590 mmol) prepared in Example 6C or obtained otherwise is dissolved in CH2Cl2 (200 mL) and prepared for oxidation by the addition of powdered 4 Å molecular sieves (ALDRICH 23,366-8, 12.00 g). 4-Methylmorpholine N-oxide (5.000 g, 42.680 mmol) and tetrapropylammonium perruthenate (VII) (0.400 g, 1.138 mmol) are then added sequentially. The reaction mixture is stirred under an inert atmosphere at ambient temperature. After about 3 days, the reaction solution is filtered through silica gel (1:1 EtOAc/CH2Cl2) and carefully purified by flash chromatography (1:1 EtOAc/CH2Cl2) to give 4-aza-5α-androstan-3,17-dione.

EXAMPLE 6E 17β-Cyclopropylamino-4-aza-5α-androstan-3-one

4-Aza-5α-androstane-3,17-dione (3.000 g, 10.366 mmol) from Example 6D is dissolved in chloroform (CHCl₃) (50 mL), cyclopropylamine (25.00 mL, 360.8 mmol) is added, and the reaction mixture is heated to reflux with stirring under a nitrogen atmosphere. After 20 h, the reaction mixture is allowed to cool to room temperature and a solution of sodium borohydride (NaBH₄; 4.000 g, 105.7 mmol) in ethanol (100 mL) is added. The reaction mixture is stirred for an additional five hours, then diluted with EtOAc (200 mL). The diluted solution is washed with water (2 x 200 mL) and brine (150 mL). The organic solution is dried over MgSO4, filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (eluting first with 1:1:2 EtOAc/CH2Cl2/hexane, then with 15:85 i-PrOH/CH2Cl2) to give the title compound in excellent purity.

PRODUCTION 7 1-Fluoro-17β-cyclopropyloxy-4-methyl-4-aza-5α-androstan-3-one PRODUCTION 7A 17β-Acetoxy-4-methyl-4-aza-5α-androstan-3-one

The 17β-acetoxy-4-methyl-4-azaandrost-5(6)-en-3-one prepared in Example 3A or otherwise obtained (20.000 g, 57.890 mmol) and 5% palladium on carbon catalyst (2.000 g, 0.940 mmol) are placed in a 500 mL Paff bottle under an inert atmosphere. Acetic acid (150 mL) is added to the reaction vessel, which is then charged with hydrogen gas to 60 p.s.i. The hydrogenation reaction takes place at 60°C with shaking. After 3 days, the reaction mixture is filtered through Celite® and concentrated to about 50 mL under reduced pressure. The concentrated product solution is poured into ice-cold water (800 mL) with stirring. The resulting precipitate is collected by filtration and recrystallized from ethanol to give the title compound.

PRODUCTION 7B 17β-Acetoxy-4-methyl-4-aza-5α-androst-1(2)-en-3-one

17β-Acetoxy-4-methyl-4-aza-5α-androstan-3-one (18.000 g, 52.098 mmol) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone prepared in Preparation 7A or obtained otherwise were dissolved in 1,4-dioxane (200 mL). Bis(trimethylsilyl)trifluoroacetamide (136.0 mL, 512.0 mmol) was carefully added to the dioxane solution while maintaining an inert atmosphere. After stirring at room temperature for 60 minutes, the reaction mixture is heated to reflux. Reflux is continued for eighteen hours, at which time the reaction mixture is allowed to cool to ambient temperature. The reaction solvent is removed under reduced pressure and the residue is redissolved in EtOAc (500 mL). The organic solution is washed with 5% sodium hydroxide solution (400 mL) and brine (400 mL), then dried over MgSO4, filtered and concentrated under reduced pressure. The reaction product is further purified by flash chromatography (1:1 EtOAc/CH2Cl2) to give the title compound.

PRODUCTION 7C 17β-Hydroxy-4-methyl-4-aza-5α-androst-1(2)-en-3-one

The 17β-acetoxy-4-methyl-4-aza-5α-androst-1(2)-en-3-one prepared in Preparation 7B (15.000 g, 43.673 mmol) was dissolved in a mixture of ethanol (100 mL) and THF (100 mL). To the ethanolic solution was added 6 M sodium hydroxide (100 mL), then the reaction mixture was stirred at room temperature for 7 h. At the end of this time, the reaction mixture was diluted with EtOAc (600 mL) and extracted with brine (3 x 400 mL). The organic phase was dried over MgSO4, filtered and evaporated to dryness to give a crude product of sufficient purity for subsequent synthesis.

MANUFACTURING 7D 17β-Cyclopropyloxy-4-methyl-4-aza-5α-androst-1(2)-en-3-one

17β-Hydroxy-4-aza-4-methyl-5α-androst-1(2)-en-3-one (12.000 g, 39.552 mmol) from Preparation 7C was dissolved in a mixture of CH2Cl2 (200 mL) and ethyl vinyl ether (200 mL, 2.091 mol). Mercury(II) acetate (Hg(OAc)2; 13.500 g, 42.362 mmol) was added to the reaction mixture, which was then heated to reflux under an inert atmosphere. After three days, the reaction solution was cooled to room temperature and acetic acid (8.00 mL, 139.7 mmol) was added. The acidified reaction mixture was stirred at room temperature for 2 h, then diluted with hexane (500 mL). The organic solution is extracted with 10% sodium hydroxide solution (300 mL) and brine (2 x 200 mL), dried over MgSO4, filtered and evaporated to dryness to give 17β-vinyloxy-4-methyl-4-azaandrost-1-en-3-one, which is used in the following synthesis without further purification.

The 17β-vinyloxy-4-methyl-4-azaandrost-1-en-3-one (8.000 g, 24.279 mmol) prepared in the previous paragraph is dissolved in a mixture of CH₂Cl₂ (100 mL) and methyl t-butyl ether (100 mL) and then quenched to 0 °C under a nitrogen atmosphere. Diethylzinc solution (1.1 M in toluene; 145 mL, 159.5 mmol) is added to the steroid solution with stirring, followed by careful dropwise addition of CH2I2 (13.00 mL, 161.4 mmol). The reaction mixture is stirred at 0 °C for 2.5 h, then allowed to warm to room temperature over 14 h. The reaction is then quenched with saturated aqueous NH4Cl solution (300 mL) and extracted with EtOAc (500 mL). The organic extract is washed with brine (2 x 300 mL), dried over MgSO4, filtered and evaporated to dryness. The crude product is purified by flash chromatography (1:1 EtOAc/CH2Cl2) to give the title compound.

PRODUCTION 7E 17β-Cyclopropyloxy-4-methyl-1-phenylthio-4-aza-5α-androstan-3-one

Sodium hydride (50% dispersion in mineral oil, 0.580 g, 12.08 mmol) is washed with hexane (3 x 25 mL) under an inert atmosphere to remove the mineral oil. Anhydrous tetrahydrofuran (THF, 50 mL) is carefully added while maintaining the inert atmosphere. A solution of thiophenol (1.230 mL, 11.98 mmol) in THF (50 mL) is slowly added dropwise over 30 min with stirring while evolving gas. At the end of the addition, the THF solution is heated to reflux for 30 min and then allowed to cool to room temperature. With stirring, a solution of 17β-cyclopropyloxy-4-methyl-4-aza-5α-androstan-1-en-3-one (4.000 g, 11.645 mmol) in THF (50 mL) is added to the thiophene oxide slurry in small portions over 20 min. Under an inert atmosphere, the reaction mixture is stirred at ambient temperature for 40 min after the addition, then heated to reflux. After refluxing for 2 h, the reaction mixture is cooled to room temperature and carefully poured into ice water (500 mL). The reaction mixture is then extracted with EtOAc (3 x 500 mL), the combined organic layers are washed with brine (2 x 250 mL), dried over MgSO4, filtered, and evaporated to dryness. Purification by flash chromatography (1:1:2 EtOAc/CH2Cl2/hexane) gives the title compound. The compound has the following structure:

EXAMPLE 7F 2-Bromo-17β-cyclopropyloxy-1-fluoro-4-methyl-4-aza-5α-androst-1-en-3-one

In a manner similar to that disclosed in R. Bohlman, Tet. Lett. 1994, 35(1), 85- 88, a solution of 17β-cyclopropyloxy-4-methyl-1-phenylthio-4-aza-5α- androstan-3-one (3.000 g, 6.613 mmol) in CH2Cl2 (100 mL) is prepared at 0 °C. N-Bromosuccinimide (2.360 g, 13.259 mmol) and diethylaminosulfur trifluoride (0.880 mL, 6.661 mmol) under an inert atmosphere are added sequentially and the reaction mixture is stirred at 0 °C for 4.5 hours, then poured into saturated aqueous NaHCO3 solution (50 mL) and extracted with CH2Cl2 (3 x 50 mL). The combined organic extracts are washed with brine (100 mL), dried over MgSO4, filtered and evaporated to dryness. The product is purified by flash chromatography (1:1 EtOAc/CH2Cl2) to give 2-bromo-17β-cyclopropyloxy-1-fluoro-4-methyl-4-aza-5α-androst-1-en-3-one.

MANUFACTURING 7G 17β-Cyclopropyloxy-1-fluoro-4-methyl-4-aza-5α-androst-1-en-3-one

In a manner similar to that disclosed in R. Bohlman, Tet. Lett. 1994, 35(1), 85-88, 2-bromo-17β-cyclopropyloxy-1-fluoro-4-methyl-4-aza-5α-androst-1-en-3-one (1.000 g, 2.270 mmol) is slurried in toluene (40 mL). Under nitrogen, tributyltin hydride (0.680 mL, 2.528 mmol) and azobisisobutyronitrile (0.0750 g, 0.457 mmol) are added to the toluene slurry and the reaction mixture is heated to 80 °C while stirring. After 3 hours, the reaction mixture is cooled to room temperature and passed through silica gel (1:1 EtOAc/CH2Cl2). The eluate is concentrated and purified by flash chromatography (1:1 EtOAc/CH2Cl2) to give the title compound.

PRODUCTION 7H 17β-Cyclopropyloxy-1-fluoro-fluoro-methyl-methyl-aza-5α-androstan-3-one

In a manner similar to that disclosed in T. Kitazume et al., J. Org. Chem. 1989, 54 (23), 5630-5632, 17β-cyclopropyloxy-1-fluoro-4-methyl-methyl-aza-5α-androst-1-en-3-one (0.250 g, 0.692 mmol) and 10% Pd/C catalyst (0.0250 g, 0.0235 mmol Pd) are charged to a flask which is then purged with nitrogen. Ethanol (10 mL) is added and the vessel is charged with hydrogen (1 atmosphere). After 30 h of ultrasonic irradiation (32 KHz, 35 W), the reaction mixture is filtered through Celite® and evaporated to dryness. The residue is purified by flash chromatography (1 : 1 EtOAc/CH₂Cl₂) to give the title compound:

PRODUCTION 8 1-Phenylsulfinyl/phenylsulfonyl-17β-cyclopropyloxy-4-methyl-4-aza-5α-androstan-3-one PRODUCTION 8A 17β-Cyclopropyloxy-1-phenylsulfinyl-methyl-methyl-4-aza-5α-androstan-3-one

17β-Cyclopropyloxy-4-methyl-1-phenylthio-4-aza-5α-androstan-3-one (3.000 g, 6.613 mmol) prepared as in Preparation 7E or obtained otherwise was dissolved in CH2Cl2 (100 mL) and quenched to -78 °C under an inert nitrogen atmosphere. 3-Chloroperoxybenzoic acid (55% in water and 3-chlorobenzoic acid; 2.250 g, 7.171 mmol) was added and the reaction mixture was stirred at -78 °C for 3 hours while maintaining the nitrogen atmosphere, after which the reaction was quenched with saturated aqueous Na2SO3 solution (100 mL). The organic layer is separated and washed with 1 M NaOH (3 x 25 mL) and brine (50 mL), dried over MgSO4, filtered and evaporated to dryness. Purify the residue by flash chromatography (1:1 EtOAc/CH2Cl2) to give the title compound.

PRODUCTION 8B 17β-Cyclopropyloxy-1-phenylsulfonyl-4-methyl-4-aza-5α-androstan-3-one

Under nitrogen, 17β-cyclopropyloxy-1-phenylsulfinyl-4-methyl-4-aza-5α-androstan-3-one (2.000 g, 4.260 mmol) is dissolved in CH2Cl2 (70 mL). 3-Chloroperoxybenzoic acid (55% in water and 3-chlorobenzoic acid; 1.500 g, 4.781 mmol) is added to the solution. After stirring under nitrogen for 16 h, the reaction is quenched with saturated aqueous Na2SO3 solution (70 mL). The organic layers are separated and washed with 1 M NaOH (3 x 25 mL) and brine (50 mL), dried over MgSO4, filtered and evaporated to dryness. Purify the residue by flash chromatography. gives the title compound which has the following formula:

PRODUCTION 9 1-Fluoro-17β-cyclopropylamino-4-methyl-4-aza-5α-androstan-3-one PRODUCTION 9A 17β-Cyclopropylamino-4-methyl-4-aza-5α-androst-1(2)-en-3-one

17β-Hydroxy-4-methyl-4-aza-5α-androst-1(2)-en-3-one (from Preparation 7C or otherwise obtained) is C17-oxidized (17-one) as in Example 6D. The C17 atom is then cycloaminated in a manner similar to that described in Example 6E to give the title compound.

PRODUCTIONS 9B, 9C, 9D, 9E 17β-Cyclopropylamino-4-methyl-1-phenylthio-4-aza-5α-androstan-3-one 2-Bromo-17β-cyclopropylamino-1-fluoro-4-methyl-4-aza-5α-androst-1(2)-en-3-one 17β-Cyclopropylamino-1-fluoro-4-methyl-4-aza-5α-androst-1(2)-en-3-an 17β-Cyclopropylamino-1-fluoro-4-methyl-4-aza-5α-androstan-3-an

17β-Cyclopropylamino-4-methyl-4-aza-5α-androst-1(2)-en-3-one is reacted in a manner similar to that described in Preparations 7E-H, respectively, to give the corresponding title compounds.

PRODUCTION 10 1-Phenylthio/phenylsulfinyl/phenylsulfonyl-4-methyl-17β-cyclopropylamino-4-aza-5α-androstan-3-one PRODUCTIONS 10A, 10B 17β-Cyclopropylamino-4-methyl-1-phenylsulfinyl-aza-5α-androstan-3-one 17β-Cyclopropylamino-4-methyl-1-phenylsulfonyl-4-aza-5α-androstan-3-one

17β-Cyclopropylamino-4-methyl-1-phenylthio-aza-5α-androstan-3-one from Preparation 9B or otherwise obtained is reacted in a manner similar to the procedures described in Preparations 8A and 8B, respectively, to afford the corresponding title compounds.

EXAMPLE 11 2-Halogen/methylthio/methylsulfinyl/methylsulfonyl-17β-cyclopropyloxy-4-aza-5α-androst-3-one EXAMPLE 11A 17β-Trimethylsilyloxy-4-aza-5α-androstan-3-one

To 17β-hydroxy-4-aza-5α-androstan-3-one prepared in Example 2B or otherwise obtained (20.000 g, 68.634 mmol) in CH2Cl2 (500 mL) is added trimethylsilyl chloride (TMSCl) (18.00 mL, 141.83 mmol) with stirring under nitrogen. Triethylamine (20.00 mL, 143.49 mmol) is added in small portions over 30 minutes. After vigorous stirring under nitrogen for 48 hours, the reaction mixture is filtered through Celite®. The filtrate is evaporated to dryness and redissolved in ether (500 mL). The ether solution is filtered through Celite® and concentrated under reduced pressure to give the title compound.

EXAMPLE 11B 17β-Hydroxy-2α-bromo-4-aza-5α-androstan-3-one

17β-Trimethylsilyloxy-4-aza-5α-androstan-3-one from Example 11A (10.000 g, 27.517 mmol) is prepared in solution with toluene (100 mL). N,N,N',N'-Tetramethylethylenediamine ((CH3)2NCH2CH2N(CH3)2, TMEDA, 12.50 mL, 82.824 mmol) is added under nitrogen. The solution is quenched to -37 °C and tetramethylsilyl iodide (TMSI, 3.95 mL, 27.76 mmol) is added dropwise. The resulting slurry is stirred for 5 minutes and then bromine (7.00 mL, 135.87 mmol) is added dropwise. With stirring, the reaction mixture is allowed to warm to 20 °C, after which it is poured into saturated aqueous sodium sulfite solution (Na₂SO₃, 100 mL). The biphasic mixture is extracted with EtOAc (100 mL) and the organic layers are combined and washed twice with Na₂SO₃ solution (100 mL) and then with brine (2 x 100 mL). The Material was then dried over MgSO4, filtered and evaporated to dryness. The residue was redissolved in THF (100 mL) and a tetrabutylammonium fluoride solution (1 M in THF; 28.00 mL, 28.00 mmol) was added to the THF solution and the mixture was then stirred at room temperature for 15 min. After this period, the reaction mixture was poured into saturated aqueous NH4Cl solution (100 mL) and extracted with EtOAc (3 x 200 mL). The organic extractions were combined, washed twice with brine (150 mL), dried over MgSO4, filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography (1:1 EtOAc/CH2Cl2) to afford the title compound in sufficient purity for the next synthesis.

EXAMPLE 11C 17β-Cyclopropyloxy-2α-bromo-4-aza-5α-androstan-3-one

17β-Hydroxy-2α-bromo-4-aza-5α-androstan-3-one is etherified and cyclopropanated to the title compound in a manner similar to that described in Preparation 7D.

EXAMPLE 11D 17β-Hydroxy-2α-iodo-4-aza-5α-androstan-3-one

The 17β-trimethylsilyloxy-4-aza-5α-androstan-3-one (10.000 g, 27.517 mmol) prepared in Example 11A or obtained otherwise is dissolved in toluene (100 mL) and N,N,N',N-tetramethylethylenediamine (TMEDA) (12.50 mL, 82.824 mmol) is added under nitrogen. The solution is quenched to -37°C and tetramethylsilyl chloride (7.35 mL, 57.91 mmol) is quickly added dropwise. The resulting slurry is stirred for 5 minutes and then iodine (10.00 g, 39.40 mmol) is added in a continuous portion. The mixture is allowed to warm to 20 °C, at which time it is poured into saturated aqueous sodium sulfite solution (Na2SO3, 100 mL). The resulting biphasic mixture is extracted with EtOAc (100 mL), the organic layers are combined and washed with saturated aqueous sodium sulfite solution (Na2SO3, 100 mL) and brine (100 mL), dried over MgSO4, filtered and evaporated to dryness. The residue is redissolved in THF (100 mL), tetrabutylammonium fluoride (1 M in THF; 28.00 mL, 28.00 mmol) is added and the mixture is stirred at room temperature for 15 min. At the end of this period, the reaction mixture was poured into saturated aqueous NH4Cl solution (100 mL) and extracted with EtOAc (3 x 200 mL). The combined organic extracts were washed with brine (2 x 150 mL), dried over MgSO4, filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography (1:1 EtOAc/CH2Cl2) to afford the title compound in a purity sufficient for subsequent syntheses.

EXAMPLE 11E 17β-Cyclopropyloxy-2α-iodo-4-aza-5α-androstan-3-one

17β-Hydroxy-2α-iodo-4-aza-5α-androstan-3-one is treated in a manner similar to the procedures described in Preparation 7D to give the title compound.

EXAMPLE 11F 17β-Cyclopropyloxy-2α-methylthio-4-aza-5α-androstan-3-one

17β-Cyclopropyloxy-2α-iodo-4-aza-5α-androstan-3-one (2.500 g, 5.470 mmol) from Example 11E or otherwise obtained is slurried in ethanol (50 mL), treated with sodium thiomethoxide, and the mixture is heated to reflux under an inert atmosphere. After 20 h, the reaction solution is allowed to cool to room temperature and poured into saturated aqueous NH4Cl solution (200 mL). The solution is extracted with EtOAc (200 mL), the organic extracts combined, washed with brine (2 x 150 mL), dried over MgSO4, filtered, and evaporated to dryness. The residue is purified by flash chromatography (1:1 EtOAc/CH2Cl2) to give the title compound in sufficient purity for subsequent syntheses. The compound has the following structure:

EXAMPLES 11G-11H 17β-Cyclopropyloxy-2α-methylsulfinyl-4-aza-5α-androstan-3-one 17β-Cyclopropyloxy-2α-methylsulfonyl-4-aza-5α-androstan-3-one

17β-Cyclopropyloxy-2α-methylthio-4-aza-5α-androstan-3-one is reacted in a manner similar to that described in Preparations 8A and 8B, respectively, to give the corresponding title compounds.

EXAMPLE 12 2α-Halogen/methylthio/methylsulfinyl/methylsulfonyl-17β-cyclopropylamino-4-aza-5α-androstan-3-one EXAMPLE 12A 17β-Cyclopropylamino-2α-iodo-4-aza-5α-androstan-3-one

The 17β-hydroxy-2α-iodo-4-aza-5α-androstan-3-one prepared in Example 11D or otherwise obtained is oxidized in a manner similar to the procedure described in Example 6D to give 2α-iodo-4-aza-5α-androstan-3,17-dione (the dione). The dione is then converted to the 17-cyclopropylimine and subsequently to the title compound in a manner similar to that described in Example 6E.

EXAMPLE 12B 17β-Cyclopropylamino-2α-methylthio-4-aza-5α-androstan-3-one 17β-Cyclopropylamino-2α-methylsulfinyl-4-aza-5α-androstan-3-one 17β-Cyclopropylamino-2α-methylsulfonyl-4-aza-5α-androstan-3-one

The above title compounds are prepared from 17β-cyclopropylamino-2α-iodo-4-aza-5α-androstan-3-one prepared in Example 12A or otherwise obtained, which is converted to the corresponding title compounds in a manner similar to that described in Examples 11F, 11G, or 11H, respectively, to afford the corresponding title compound.

EXAMPLE 12C 17β-Cyclopropylamino-2α-bromo-4-aza-5α-androstan-3-one

The 17β-hydroxy-2α-bromo-4-aza-5α-androstan-3-one prepared in Example 11B or otherwise obtained is oxidized in a manner similar to that described in Example 6D to give 2α-bromo-4-aza-5α-androstan-3,17-dione (the dione). The dione is then converted to the 17-cyclopropylimine and then to the title compound in a manner similar to that described in Example 6E.

EXAMPLE 13 17-Cyclopropyloxy-4-methyl-4-aza-5α-androstan-3-one EXAMPLE 13A 17β-Hydroxy-4-methyl-4-aza-5α-androstan-3-one

The 17β-hydroxy-4-methyl-4-aza-androst-5(6)-en-3-one prepared in Example 3B or otherwise obtained is hydrogenated under conditions similar to those described in Example 6C to give the title compound.

EXAMPLE 13B 17β-Cyclopropyloxy-4-methyl-4-aza-5α-androstan-3-one

The 4-methyl-17β-hydroxy-4-aza-5α-androstan-3-one prepared in Example 13A or otherwise obtained is etherified and cyclopropanated as described in Preparation 7D to give the title compound.

EXAMPLE 13C 17β-Cyclopropylamino-4-methyl-4-aza-5α-androstan-3-one

The 17β-hydroxy-4-methyl-4-aza-5α-androstan-3-one prepared in Example 13A or otherwise obtained is oxidized to 4-methyl-4-aza-5α-androst-3,17-dione (the dione) as described in Example 6D. The dione is then cycloaminated as described in Example 6E to give the title compound.

EXAMPLE 14 2α-Halogen/methylthio/methylsulfinyl/methylsulfonyl-4-methyl-17β-cyclopropyloxy-4-aza-5α-androstan-3-one EXAMPLE 14A 4-Methyl-17β-trimethylsilyloxy-4-aza-5α-androstan-3-one

The 17β-hydroxy-4-methyl-4-aza-5α-androstan-3-one prepared in Example 13A or otherwise obtained is silylated as described in Example 11A to give the title compound.

EXAMPLE 14B 2α-Bromo-17β-cyclopropyloxy-4-methyl-4-aza-5α-androstan-3-one

4-Methyl-17β-trimethylsilyloxy-4-aza-5α-androstan-3-one from Example 14A or otherwise obtained is brominated and cyclopropanated in a manner similar to the procedure described in Examples 11B or 11C, respectively, to give the title compound.

EXAMPLE 14C 2α-iodo-17β-cyclopropyloxy-4-methyl-4-aza-5α-androstan-3-one

4-Methyl-17β-trimethylsilyloxy-4-aza-5α-androstan-3-one from Example 14A or otherwise obtained is iodinated and cyclopropanated in a similar manner as in Examples 11D or 11E, respectively, to give the title compound.

EXAMPLES 14D, 14E AND 14F 17β-Cyclopropyloxy-4-methyl-2α-methylthio-4-aza-5α-androstan-3-one 17β-Cyclopropyloxy-4-methyl-2α-methylsulfinyl-4-aza-5α-androstan-3-one 17β-Cyclopropyloxy-4-methyl-2α-methylsulfonyl-4-aza-5α-androstan-3-one

2α-iodo-17β-cyclopropyloxy-4-methyl-4-aza-5α-androstan-3-one from Example 14C or otherwise as described in Examples 11F, 11G or 11H, respectively, to afford the above corresponding title compounds.

EXAMPLE 15 2α-Halogen/methylthio/methylsulfinyl/methylsulfonyl-4-methyl-17β-cyclopropylamino-4-aza-5α-androstan-3-one EXAMPLE 15A 2α-iodo-17β-cyclopropylamino-4-methyl-4-aza-5α-androstan-3-one

2α-Iodo-17β-hydroxy-4-methyl-4-aza-5α-androstan-3-one is oxidized and cycloaminated as in Examples 6D and 6E, respectively, to give the title compound.

EXAMPLES 15B, 15C AND 15D 17β-Cyclopropylamino-4-methyl-2α-methylthio-4-aza-5α-androstan-3-one 17β-Cyclopropylamino-4-methyl-2α-methylsulfinyl-4-aza-5α-androstan-3-one 17β-Cyclopropylamino-4-methyl-2α-methylsulfonyl-4-aza-5α-androstan-3-one

2α-Iodo-17β-cyclopropylamino-4-methyl-4-aza-5α-androstan-3-one is reacted in a manner similar to the procedures described in Examples 11F, 11G, and 11H, respectively, to give the corresponding title compounds above.

EXAMPLE 15E 2α-Bromo-17β-cyclopropylamino-4-methyl-4-aza-5α-androstan-3-one

The 2α-bromo-17β-trimethylsilyloxy-4-methyl-4-aza-5α-androstan-3-one prepared as in Example 14B is hydrolyzed as in Example 11B to give 2α-bromo-17β-hydroxy-4-methyl-4-aza-5α-androstan-3-one (17-alcohol). The 17-alcohol is then oxidized and cycloaminated as in Examples 6D and 6E, respectively, to give the title compound.

EXAMPLE 16 7β-Ethyl-17β-cyclopropyloxy-4-azaandrost-5(6)-en-3-one EXAMPLE 16A 3β-Hydroxy-17β-tert-butyldimethylsilyloxyandrost-5(6)-ene-3β-acetate

3β,17β-Dihydroxyandrost-5(6)-ene-3-acetate (50.000 g, 150.38 mmol), obtained by treating 3β-acetoxyandrost-5(6)-en-17-one (available from ALDRICH, 39,008-9) with sodium borohydride, was dissolved in CH2Cl2 (225 mL) with t-butyldimethylsilyl chloride (30.000 g, 199.0 mmol) under nitrogen. 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU, 33.00 mL, 220.7 mmol) was added dropwise to the CH2Cl2 solution while maintaining the temperature below reflux with an ice bath. After stirring for 72 hours at room temperature, the reaction solution is diluted with EtOAc (800 mL) and extracted with saturated aqueous NH4Cl solution (3 x 200 mL), brine (200 mL), saturated aqueous NaHCO3 solution (200 mL) and once again with brine (200 mL). The combined organic layers are dried over MgSO4, filtered and concentrated under reduced pressure to give the title compound in a purity sufficient for subsequent synthesis.

EXAMPLE 16B 3β-Acetoxy-17β-tert-butyldimethylsilyloxyandrost-5(6)-en-7-one

In a manner similar to the procedure described in Pinto et al., Chem. Pharm. Bull., 1988, 36(12), 4689-4692 and T. Kutney, C. Gletus, Steroids 1966, 7(1), 67-78, 3β-hydroxy-17β-tert-butyldimethylsilyloxyandrost-5(6)-ene-3-acetate (60.000 g, 134.3 mmol) is dissolved in carbon tetrachloride (CCl4, 250 mL). Acetic anhydride (38.00 mL, 402.7 mmol), acetic acid (100.0 mL, 1746.8 mmol) and t-butyl chromate are added sequentially to the CCl4 solution, which is then heated to reflux temperature under an inert atmosphere. After 29 hours, the reaction mixture is cooled to 0 °C and poured into a solution of oxalic acid (70.000 g, 777.4 mmol) in water (700 mL) with stirring. An additional portion of oxalic acid (54.000 g, 599.7 mmol) is added to the biphasic mixture and stirring is continued for 4 hours while allowing the solution to warm to room temperature. Water (750 mL) is added and the biphasic mixture is extracted three times with CH2Cl2 (800 mL). The combined organic phases are washed with water (2 x 1000 ml), dried over MgSO4, filtered and evaporated to dryness. The crude product is recrystallized from ethanol to give the title compound in sufficient purity for subsequent synthesis.

EXAMPLE 16C 17β-t-Butyldimethylsilyloxy-7-ethylandrost-5-en-3β,7-diol

3β-Acetoxy-17β-t-butyldimethylsilyloxyandrost-5(6)-en-7-one (40.000 g, 86.821 mmol) is dissolved in THF (500 mL) under an inert atmosphere. A solution of ethylmagnesium chloride (2.0 M in THF; 90.0 mL, 180.0 mmol) is added to the steroid solution in portions over 60 min. At the end of the addition, the reaction mixture is stirred at room temperature for six hours and then heated to reflux. After 16 hours at reflux, the Grignard reaction mixture is allowed to cool to room temperature and then poured into saturated aqueous NH4Cl solution (500 mL). The reaction product is extracted into EtOAc (3 x 500 mL) and the combined extracts are washed with saturated aqueous NaHCO3 solution (3 x 500 mL) and brine (500 mL), dried over MgSO4, filtered and concentrated to give the crude product as a mixture of diastereomers which is carried to the next step without further purification.

EXAMPLE 16D 7-Ethyl-17β-tert-butyldimethylsilyloxyandrost-4,6-dien-3-one

In a manner similar to the procedure described in J. Eastham & R. Teranishi, Org. Synth. Coll. Vol. IV 1963, 192-195, and C. Djerassi, Org. React., 1951, 6, 207-272, 17β-t-butyldimethylsilyloxy-7-ethylandrost-5-ene-3β,7-diol (35.000 g, 71.324 mmol) prepared in Example 16C is dissolved in a mixture of toluene (700 mL) and cyclohexanone (200 mL). To the toluene solution is added dropwise a solution of aluminum isopropoxide (10.000 g, 48.960 mmol) in toluene (150 mL). The reaction mixture is heated to reflux under an inert atmosphere. After about thirty minutes, or when the reaction volume has condensed to about half the original volume, saturated aqueous potassium sodium tartrate tetrahydrate solution [KO2-CH(OH)CH(OH)CO2Na·4H2O, 150 mL] is added and the reaction mixture is refluxed with vigorous stirring for 30 minutes. At the end of this time, the biphasic solution is allowed to cool to room temperature and the phases are separated. The aqueous phase is extracted three times with CH2Cl2 and the combined organic phases are washed with brine (2 x 100 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The residue is recrystallized from ethanol to give the title compound.

EXAMPLE 16E 7β-Ethyl-17β-tert-butyldimethylsilyloxyandrost-5-en-3-one

In a manner similar to the procedures described in S. Crabtree et al., Org. Synth. 1991, 70, 256-264, D. Caine et al., Org. Synth. Coll. Vol. VI, 1988, 51-55 and D. Caine, Org. React., 1976, 23, 1-258, small pieces of lithium metal (0.950 g, 136.9 mmol) are added to dry liquid ammonia (3.00 mL) at -78 °C under an inert atmosphere. After stirring at -78°C for 20 min or until the lithium is dissolved, a solution of 7β-ethyl-17β-tert-butyldimethylsilyloxyandrost-4,6-dien-3-one from Example 16D or otherwise obtained in a mixture of t-butanol (5.70 mL, 60.45 mmol) and toluene (50 mL) is added dropwise to the blue ammonia solution over 60 min while maintaining the temperature at -78°C. After the addition, the ammonia solution is stirred for an additional 15 min, after which a quench with solid NH₄Cl (15.00 g, 280.4 mmol) is applied. The ammonia is allowed to evaporate from the reaction mixture under a dry nitrogen stream while allowing the reaction vessel to warm to room temperature. The remaining toluene slurry is diluted with EtOAc (300 mL), washed with water (200 mL), washed again with brine (2 x 200 mL), dried over MgSO4, filtered and evaporated to dryness. The residue is recrystallized from ethanol to give the title compound.

EXAMPLE 16F 7β-Ethyl-17β-hydroxyandrost-4-en-3-one

7β-Ethyl-17β-tert-butyldimethylsilyloxyandrost-5-en-3-one (15.000 g, 34.824 mmol) from Example 16E or otherwise obtained is dissolved in THF (100 mL). To the solution is added 1,8-diazabicyclo[5.4.0]undec-7-ene (1.20 mL, 8.024 mmol) and the reaction mixture is heated to reflux under nitrogen. After 1 hour, the reaction mixture is cooled to room temperature and tetrabutylammonium fluoride (1.0 M in THF; 36.0 mL, 36.00 mmol) is added. The desilylation reaction is allowed to proceed for 90 min at room temperature with stirring. At the end of this time, the reaction mixture is diluted with EtOAc (500 mL) and washed with saturated aqueous NH4Cl solution (3 x 200 mL) and brine (200 mL), then dried over MgSO4, filtered and evaporated to dryness. The residue is recrystallized from ethanol to give the title compound.

EXAMPLE 16G 7β-Ethyl-17β-hydroxy-4-azaandrost-5-en-3-one

7β-Ethyl-17β-hydroxyandrost-4-en-3-one (10.000 g, 34.914 mmol) is converted to 17β-hydroxy-7β-ethyl-5-oxo-4-nor-3,5-seco-androstane-3-carboxylic acid (seco acid) as in Example 6A. The seco acid is then converted to the β-lactam similar to the procedure described in Example 6B. The β-lactam is then hydrolyzed to 7β-ethyl-17β-hydroxy-4-azaandrosten-3-one in a similar manner as described in Preparation 7C.

EXAMPLE 16H 17β-Cyclopropyloxy-7β-ethyl-4-aza-5α-androst-5-en-3-one

The 7β-ethyl-17β-hydroxy-4-azaandrost-5-en-3-one prepared in Example 16G above is etherified and cyclopropanated as described in Preparation 7D to give the title compound.

EXAMPLE 17 17β-Cyclopropyl(oxy/amino)-7β-ethyl-4-aza-5α-androst-[1(2)en/an]-3-one EXAMPLE 17A 17β-Acetoxy-7β-ethyl-4-aza-5α-androstan-3-one

The 17β-acetoxy-7β-ethyl-4-aza-androst-5-en-3-one prepared in the second paragraph of Example 16G is hydrogenated in a similar manner to Example 6C to give the title compound.

EXAMPLE 17B 17β-Cyclopropyloxy-7β-ethyl-4-aza-5α-androstan-3-one

17β-Acetoxy-7β-ethyl-4-aza-5α-androstan-3-one from Example 17A is hydrolyzed as described in Preparation 7C, then etherified and cyclopropanated as described in Preparation 7D to give the title compound.

EXAMPLE 17C 17β-Hydroxy-7β-ethyl-4-aza-5α-androst-1(2)-en-3-one

17β-Acetoxy-7β-ethyl-4-aza-5α-androstan-3-one from Example 17A is dehydrated as in Preparation 7B to give 17β-acetoxy-7β-ethyl-4-aza-5α-androst-1(2)-en-3-one which is then hydrolyzed as in Preparation 7C to give the title compound.

EXAMPLE 17D 17β-Cyclopropyloxy-7β-ethyl-4-aza-5α-androst-1(2)-en-3-one

17β-Hydroxy-7β-ethyl-4-aza-5α-androst-1(2)-en-3-one from Example 17C is etherified and cyclopropanated as described in Preparation 7D to give the title compound.

EXAMPLE 17E 17β-Cyclopropylamino-7β-ethyl-4-aza-5α-androst-1-en-3-one

17β-Hydroxy-7β-ethyl-4-aza-5α-androst-1(2)-en-3-one from Example 17C is C₁₇-oxidized as described in Example 6D, then cycloaminated as described in Example 6E to give the title compound.

EXAMPLE 17F 17β-Cyclopropylamino-7β-ethyl-4-aza-5α-androstan-3-one

17β-Acetoxy-7β-ethyl-5α-androstan-3-one from Example 17A is hydrolyzed as in Preparation 7C, then C17-oxidized as in Example 6D, then cycloaminated as in Example 6E to give the title compound.

EXAMPLE 18 17β-Cyclopropyloxy-4-azaandrost-3,7-dione EXAMPLE 18A 7,7-Ethylenedioxy-17β-t-butyldimethylsilyloxyandrost-5-en-3-ol-3-acetate

In a manner similar to that described in T. Tsunoda et al., Tet. Lett., 1980, 21(14), 1357-1358, Hwu et al., J Org. Chem., 1987, 52(2), 188-191, a solution of 3β-acetoxy-17β-t-butyldimethylsilyloxyandrost-5-en-7-one (40.000 g, 86.821 mmol) obtained from Example 16B or otherwise is prepared and 1,2-bis(trimethylsiloxy)ethane (23.00 mL, 93.81 mmol) in CH2Cl2 (300 mL) at -78°C. Trimethylsilyl trifluoromethanesulfonate (0.20 mL, 1.035 mmol) is added under an inert atmosphere, the reaction mixture is stirred for 3 hours, pyridine (2.00 mL, 24.73 mmol) is added and the mixture is allowed to warm to room temperature with stirring. The organic solution is poured into saturated aqueous NaHCO3 solution (300 mL) and the resulting biphasic solution is extracted with EtOAc (3 x 300 mL). The combined organic extracts are washed with brine (200 mL), dried over MgSO4, filtered and evaporated to dryness. The residue is purified by flash chromatography (1:4 EtOAc/hexane) to give 7,7-ethylenedioxy-17β-t-butyldimethylsilyloxyandrost-5-en-3-ol-3-acetate.

EXAMPLE 18B 17β-Hydroxy-4-azaandrost-5-en-3,7-dione

7,7-Ethylenedioxy-17β-t-butyldimethylsilyloxyandrost-5-en-3-ol-3-acetate from Example 18A is hydrolyzed in a similar manner as described in Preparation 7C to 7,7-ethylenedioxy-17β-t-butyldimethylsilyloxyandrost-5-en-3-ol, which is oxidized by the procedure described in Preparation 16D to 7,7-ethylenedioxy-17β-t-butyldimethylsilyloxyandrost-5-en-3-one, which is then isomerized and desilylated in a manner described in Example 16F to give 7,7-ethylenedioxy-17β-hydroxyandrost-4-en-3-one.

7,7-Ethylenedioxy-17β-hydroxyandrost-4-en-3-one is oxidized to 7,7-ethylenedioxy-17β-hydroxy-5-oxo-4-nor-3,5-seco-androstane-3-carboxylic acid (seco acid) as described in Example 6A. The seco acid is converted to 7,7-ethylenedioxy-17β-hydroxy-4-azaandrost-5-en-3,7-dione (β-lactam) in a manner similar to that described in Example 6B. The β-lactam is hydrolyzed under conditions similar to those described in Preparation 7C to give 17β-hydroxy-4-azaandrost-5-ene-3,7-dione.

EXAMPLE 18C 17β-Cyclopropyloxy-4-azaandrost-5-en-3,7-dione

17β-Hydroxy-4-azaandrost-5-ene-3,7-dione from Example 18B or otherwise obtained is cyclopropanated in a manner similar to that described in Preparation 7D to give the title compound.

EXAMPLE 18D 17β-Cyclopropyloxy-4-aza-5α-androstane-3,7-dione

17β-Cyclopropyloxy-4-azaandrost-5-ene-3,7-dione from Example 18C or otherwise is hydrogenated under similar conditions as described in Preparation 7H to give the title compound.

EXAMPLE 19 17β-Cyclopropylamino-4-azaandrost-5-en-3,7-dione EXAMPLE 19A 17β-Trimethylsilyloxy-4-trimethylsilyl-4-azaandrost-5-en-3,7-dione

17β-Hydroxy-4-azaandrost-5-ene-3,7-dione (40.000 g, 131.84 mmol) prepared in Example 18B or otherwise obtained is prepared in solution at -78°C in dry THF (300 mL). Lithium diisopropylamide solution (2.0 M in heptane/THF/ethylbenzene; 135.0 mL, 270.0 mmol) is added under an inert atmosphere and the deprotonation reaction takes place while stirring at -78°C for 60 minutes. Trimethylsilyl chloride (35.00 mL, 275.8 mmol) is added and stirring is continued for an additional 15 minutes and then the solution is allowed to warm to room temperature. After stirring for an additional 2 hours, the reaction mixture is filtered through Cellte® and the filtrate is evaporated to dryness. The residue is redissolved in ether (500 ml) and the resulting slurry is filtered again through Celite®. The solvent is evaporated to give the title compound.

EXAMPLE 19B 7-t-Butyldimethylsilyloxy-17β-cyclopropylamino-4-azaandrost-5,7-dien-3-one

By the procedure described in J. W. Hart et al., J. Chem. Soc. Chem. Commun. 1979, 156-157, 17β-trimethylsilyloxy-4-trimethylsilyl-4-azaandrost-5-ene-3,7-dione (40.000 g, 92.644 mmol) prepared in Example 19A is prepared at -78°C in a solution in a mixture of THF (300 mL) and hexamethylphosphoramide (30.0 mL, 172.4 mmol) and is reacted with lithium diisopropylamide (2.0 M in heptane/THF/ethylbenzene; 50.0 mL, 100.0 mmol) under an inert atmosphere. Deprotonation is allowed to proceed for 60 minutes with stirring at -78°C. A solution of t-butyldimethylsilyl chloride (17.00 g, 112.8 mmol) in THF (50 mL) is added to the steroid solution via cannula and the reaction mixture is stirred for an additional 30 min and then allowed to warm to room temperature. After stirring for 20 h at ambient temperature, the reaction is quenched by carefully adding saturated aqueous NH4Cl solution (300 mL). The biphasic mixture is extracted with EtOAc (3 x 400 mL) and the combined organic layers are washed with water (3 x 300 mL) and brine (300 mL), dried over MgSO4 and dried over 150 mL. dried, filtered and concentrated to give 7-(t-butyldimethylsilyloxy)-4- trimethylsilyloxy-17β-trimethylsilyloxy-4-azaandrost-5,7-dien-3-one (the residue).

The residue is taken up in THF (200 mL) and treated with a mixture of acetic acid (200 mL) and water (25 mL). The reaction mixture is stirred for 60 min at ambient temperature, then diluted with EtOAc (800 mL) and washed with brine (2 x 300 mL), water (4 x 300 mL), saturated NaHCO3 solution (2 x 300 mL) and brine (300 mL). The organic solution is dried over MgSO4, filtered and concentrated. The residue is purified by flash chromatography (1:1:2 EtOAc/CH2Cl2/hexane) to give 7-(t-butyldimethylsilyloxy)-17β-hydroxy-4-azaandrost-5,7-dien-3-one (17-alcohol).

The 17-alcohol is oxidized to 7-(t-butyldimethylsilyloxy)-4-azaandrost-5,7-dien-3,17-dione using the conditions described in Example 6D, which is then converted to 17β-cyclopropylamino-7-(t-butyldimethylsilyloxy)-4-azaandrost-5,7-dien-3-one under conditions similar to those described in Example 6E.

EXAMPLE 19C 17β-Cyclopropylamino-4-azaandrost-5-en-3,7-dione

17β-Cyclopropylamino-7-(t-butyldimethylsilyloxy)-4-azaandrost-5,7-dien-3-one (5.000 g, 10.947 mmol) prepared in Example 19B or obtained otherwise is dissolved in TEF (50 mL) under nitrogen and to this solution is added tetrabutylammonium fluoride (1.0 M in THF; 12.00 mL, 12.00 mmol). The reaction mixture is stirred at ambient temperature for 90 min, then quenched with water (200 mL). The crude product solution is extracted with EtOAc (2 x 200 mL) and the combined organic extracts are washed with brine (3 x 200 mL). The organic solution is dried over MgSO4, filtered and concentrated. The residue is purified by flash chromatography (1:1 EtOAc/CH2Cl2) to give the title compound.

EXAMPLE 19D 17β-Cyclopropylamino-4-aza-5α-androstane-3,7-dione

17β-Cyclopropylamino-4-azaandrost-5-ene-3,7-dione from Example 19C or otherwise obtained is hydrogenated in a manner similar to that described in Preparation 7H to give the title compound.

EXAMPLE 20 4-Aza-17β-hydroxy-7-oxo-androstene EXAMPLE 20A 17β-Cyclopropyloxy-17β-hydroxy-4-azaandrost-5-en-3-one

The 17β-cyclopropyloxy-4-azaandrost-5-ene-3,7-dione (1.000 g, 2.9115 mmol) prepared in Example 18C or obtained otherwise is dissolved in a mixture of THF (25 mL) and ethanol (25 mL) and added to a solution of sodium borohydride (NaBH4; 1.000 g, 26.43 mmol) in ethanol (50 mL). The reaction mixture is stirred at ambient temperature for 5 hours. At the end of this time, the reaction solution is diluted with EtOAc (100 mL) and washed with brine (NaCl; 3 x 100 mL). The organic solution is dried over MgSO4. dried, filtered, concentrated and purified by flash chromatography (1:1 EtOAc/CH2Cl2) to give the title compound.

EXAMPLES 20B, 20C AND 20D 17β-Cyclopropyloxy-7β-hydroxy-4-aza-5α-androstan-3-one 17β-Cyclopropylamino-7β-hydroxy-4-azaandrost-5-en-3-one 17β-Cyclopropylamino-7β-hydroxy-4-aza-5α-androstan-3-one

The above 7-hydroxysteroids are obtained by selectively reducing the C7 carbon atom of the corresponding 7-oxo compounds, 17β-cyclopropyloxy-4-aza-5α-androstane-3,7-dione, 17β-cyclopropylamino-4-azaandrost-5-ene-3,7-dione and 17β-cyclopropylamino-4-aza-5α-androstane-3,7-dione, obtained in Examples 18D, 19C and 19D, respectively, by the method described in Example 20A.

PRODUCTION 21 4-Aza-7β-carboxymethylandrostane PRODUCTION 21A 7β-Ethoxycarbonylmethyl-17β-hydroxy-4-aza-5α-androstan-3-one

Prepare a solution of sodium hydride (50% in mineral oil; 7.000 g, 145.8 mmol) in THF (200 mL) and wash the solution with hexane (3 x 30 mL). Carefully add triethylphosphonoacetate (28.7 mL, 144.7 mmol) dropwise under nitrogen, allowing the anion to form over 60 min. Add a solution of 17β-trimethylsilyloxy-4-trimethylsilylandrost-5-ene-3,7-dione (32.406 g, 72.374 mmol) in THF, followed by stirring at room temperature under nitrogen for 6 h. After this time, EtOAc (500 mL) is added and the reaction mixture is washed with brine (3 x 400 mL), dried over MgSO4 and dried over 1 h. dried, filtered and concentrated under reduced pressure to give 17β-trimethylsilyloxy-4-trimethylsilyl-7-ethoxycarbonylmethylene-4-azaandrost-5- en-3-one (dienidisilane).

The diene disilane prepared above is redissolved in THF (300 mL) and treated with tetrabutylammonium fluoride (1.0 M in THF; 35.0 mL, 35.0 mmol). The desilylation reaction takes place over 20 min with stirring at ambient temperature. The reaction mixture is then diluted with EtOAc (500 mL) and washed with brine (3 x 400 mL). The crude product solution is dried over MgSO4, filtered, and evaporated to dryness. The crude product is purified by flash chromatography (1:1 EtOAc/CH2Cl2) to afford 17β-hydroxy-4-trimethylsilyl-7-ethyloxycarbonylmethylene-4-azaandrost-5-en-3-one as a mixture of E and Z isomers (diolefin). The diolefin is then hydrogenated in a manner similar to that described in Preparation 7H to give 7β-ethoxycarbonylmethyl-17β-hydroxy-4-aza-5α-androstan-3-one. The Compound has the following structure:

PRODUCTION 21B 17β-Cyclopropyloxy-7β-ethoxycarbonylmethyl-4-aza-5α-androstan-3-one

7β-Ethoxycarbonylmethyl-17β-hydroxy-aza-5α-androstan-3-one from Preparation 21A is etherified and cyclopropanated in a manner similar to that described in Preparation 7D to give the title compound.

PRODUCTION 21C 17β-Cyclopropyloxy-4-aza-3-oxo-5α-androstane-7β-ethanoic acid

In a manner similar to that described in Preparation 7C, 17β-cyclopropyloxy-7β-ethoxycarbonylmethyl-4-aza-5α-androstan-3-one is selectively hydrolyzed at the C7 position to give the title compound.

EXAMPLE 22 17β-Cyclopropyloxy-7β-ethoxycarbonyl/carboxy-aza-5α-androstan-3-one EXAMPLE 22A 17β-Hydroxy-7β-(diphenylhydroxymethyl)-4-aza-5α-androstan-3-one

The 17β-hydroxy-7β-ethoxycarbonylmethyl-4-aza-5α-androstan-3-one prepared in Preparation 21A (10.000 g, 26.489 mmol) was dissolved in dry THF (200 mL) and to this solution was carefully added phenylmagnesium chloride (2.0 M in THF; 55.0 mL, 110.0 mmol) with vigorous stirring under nitrogen while maintaining the reaction solution below reflux temperature with an ice bath. The reaction mixture was stirred at room temperature for 3 hours after addition of the Grignard reagent, then heated to reflux with vigorous stirring under nitrogen for 16 hours. The reaction mixture was allowed to cool to ambient temperature and then quenched by careful addition of saturated aqueous NH4Cl solution (300 mL). The crude product slurry is extracted with EtOAc (3 x 300 mL). The combined organic extracts are washed with brine (200 mL), dried over MgSO4, filtered and evaporated to dryness. The residue is purified by flash chromatography (15:85 isopropyl alcohol/CH2Cl2) to give the title compound.

EXAMPLE 22B 3,17-Dioxo-4-aza-5α-androstane-7β-carboxylic acid

In a manner analogous to that described in B. Riegel et al., Org. Synth. Coll. Vol. 3, 1955, 234-236 and C. S. Subramaniam et al., Synthesis, 1978, 468-469, 17β-hydroxy-7β-(diphenylhydroxymethyl)-4-aza-5α-androstan-3-one (8.000 g, 16.890 mmol) prepared in Example 22A is dissolved in a mixture of CH2Cl2 (60 mL) and acetic acid (60 mL), then treated with a solution of chromium trioxide (chromic acid, CrO3; 8.500 g, 85.01 mmol) in a mixture of water (6.0 mL) and acetic acid (40 mL) at room temperature. The reaction mixture is stirred for 20 minutes and then acetic anhydride (34.0 mL, 360.3 mmol) is added. The reaction solution is heated to gentle reflux with stirring. After refluxing for 20 minutes, the reaction is quenched by the careful addition of methanol (50 mL), then cooled to room temperature and concentrated to about 75 mL. The concentrated solution is poured into ice water (500 mL) and the resulting precipitate is collected by filtration. The solid is recrystallized from ethanol to give the title compound.

EXAMPLE 22C 7β-Ethoxycarbonyl-4-aza-5α-androstane-3,17-dione

In a manner analogous to the procedure described by B. Neises and W. Steglich, Org. Synth. 1984, 63, 183-187, 3,17-dioxo-4-aza-5α-androstane-7β-carboxylic acid (3.000 g, 8.998 mmol) and 4-(dimethylamino)pyridine (1.100 g, 9.004 mmol) prepared in Example 22B are dissolved in a mixture of CH2Cl2 (50 mL) and ethanol (50 mL) under nitrogen. 1,3-Dicyclohexylcarbodiimide (2.000 g, 9.693 mmol) is added to the steroid solution and the reaction mixture is stirred at room temperature for 3 hours. The reaction solution is then filtered through Celite® to remove any dicyclohexylurea that may precipitate. The filtrate is evaporated to dryness and purified by flash chromatography (1:1 EtOAc/CH2Cl2) to give the title compound. The compound has the following formula:

EXAMPLE 22D 17β-Cyclopropyloxy-7β-ethoxycarbonyl-4-aza-5α-androstan-3-one

In a manner analogous to the procedure described in Example 20A, 7β-ethoxycarbonyl-4-aza-5α-androstan-3,17-dione from Example 22C or otherwise obtained is reduced to 17β-hydroxy-7β-ethoxycarbonyl-4-aza-5α-androstan-3-one, which is then etherified and cyclopropanated by the procedure of Preparation 7D to give the title compound.

EXAMPLE 22E 17β-Cyclopropyloxy-3-oxo-4-aza-5α-androstane-7β-carboxylic acid

17β-Cyclopropyloxy-7β-ethoxycarbonyl-4-aza-5α-androstan-3-one from Example 22D or otherwise obtained is hydrolyzed by the method of Preparation 7C to give the title compound.

EXAMPLE 23A 17β-Cyclopropyloxy-7β-propyloxycarbonyl-4-azaandrost-5-en-3-one

In a manner analogous to the procedure described in H. Baer et al., Can. J. Chem. 1991, 69, 1563-1574, 17β-cyclopropyloxy-7β-hydroxy-4-azaandrost-5-en-3-one (1.000 g, 2.895 mmol) from Example 20A or otherwise obtained is dissolved in pyridine (20 mL). Propionic anhydride (20.0 mL, 156.0 mmol) is added and the mixture is stirred at room temperature under nitrogen for 12 hours. At the end of the reaction time, the solution is acidified with CH₂Cl₂. (150 mL) and washed with water (2 x 100 mL), saturated aqueous NaHCO3 solution (2 x 100 mL), water (100 mL) and brine (100 mL). The organic phase is then dried over MgSO4, filtered and evaporated to dryness. The residue is purified by flash chromatography (1:1 EtOAc/CH2Cl2) to give the title compound.

EXAMPLE 23B 17β-Cyclopropyloxy-7β-propyloxycarbonyl-4-aza-5α-androstan-3-one

In a manner analogous to the procedure described in Example 23A, 17β-cyclopropyloxy-7β-hydroxy-4-aza-5α-androstan-3-one from Example 20B or otherwise obtained is transesterified to the title compound.

EXAMPLE 24A 17β-tert-butyldimethylsilyloxy-7β-hydroxymethyl-4-tert-butyldimethylsilyl-4-aza-5α-androstan-3-one

The 7β-ethoxycarbonyl-4-aza-5α-androstan-3,17-dione prepared in Example 22C or otherwise obtained is reduced similarly to Example 20A to give 7β-ethoxycarbonyl-17β-hydroxy-4-aza-5α-androstan-3-one (17-alcohol). The 17-alcohol is then silylated as in Example 16A to give 17β-tert-butyldimethylsilyloxy-7β-ethoxycarbonyl-4-tert-butyldimethylsilyl-4-aza-5α-androstan-3-one (protected alcohol).

In a manner analogous to the procedure described in R. W. Jeanloz & E. Walker, Carbohydrate Res. 1967, 4, 504, and E. Walker, Chem. Soc. Rev., 1976, 5, 23-50, the protected alcohol synthesized above (3.000 g, 5.0674 mmol) is dissolved in THF (50 mL) and a lithium borohydride solution (2.0 M in THF; 5.50 mL, 11.0 mmol) is added under an inert atmosphere. After stirring for 3 h at room temperature, the reaction mixture is diluted with EtOAc (200 mL) and washed with brine (3 x 100 mL). The organic solution is dried over MgSO4, filtered, and evaporated to dryness. The residue is purified by flash chromatography (1:1:2 EtOAc/CH2Cl2/hexane) to give 17β-tert-butyldimethylsilyloxy-7β-hydroxymethyl-4-tert-butyldimethylsilyl-4-aza-5α-androstan-3-one.

EXAMPLE 24B 17β-t-Butyldimethylsilyloxy-4-t-butyldimethylsilyl-3-oxo-4-aza-5α-androstane-7β-carbaldehyde

In a manner analogous to the procedure described in T. Inokuchi et al., J. Org. Chem. 1990, 55, 462-466, 17β-tert-butyldimethylsilyloxy-7β-hydroxymethyl-4-tert-butyldimethylsilyl-4-aza-5α-androstan-3-one (2.000 g, 3.6365 mmol) prepared in Example 24A and 4-hydroxy-2,2,6,6-tetramethylpiperidinyloxybenzoate (4-hydroxy-TEMPO-benzoate; 0.0250 g, 0.0905 mmol) were dissolved in CH2Cl2 (70 mL) and combined with saturated NaHCO3 solution (120 mL). The biphasic solution is quenched to 0 °C and sodium bromite (NaBrO2, 1.700 g, 12.602 mmol) is added with vigorous stirring. After the bromite addition, the reaction mixture is allowed to warm to room temperature and stirred for an additional 3 hours. The reaction is quenched by dropwise addition of ethanol (1.00 mL, 17.04 mmol) and then the layers are separated. The aqueous layer is extracted with CH2Cl2 (2 x 50 mL) and the combined organic layers are washed with brine (2 x 75 mL). The organic solution is dried over MgSO4, filtered and evaporated to dryness. The residue is purified by flash chromatography (1:1:2 EtOAc/CH2Cl2/hexanes) to give the title compound.

EXAMPLE 24C 7β-(1-Hydroxypropyl)-17β-t-butyldimethylsilyloxy-4-t-butyldimethylsilyl-4-aza-5α-androstan-3-one

In a manner analogous to the procedure described in Y. Yamamoto & J. Tamada, J. Am. Chem. Soc. 1987, 109, 4395-4396, the 17β-t-butyldimethylsilyloxy-4-t-butyldimethylsilyl-3-oxo-4-aza-5α-androstane-7β-carbaldehyde (1.000 g, 1.8250 mmol) prepared in Example 24B or otherwise obtained is dissolved in CH2Cl2 and cooled to -78°C. A solution of titanium(IV) chloride (TiCl4; 1.0 M in CH2Cl2; 2.20 mL, 2.20 mmol) and tetraethyl lead (Et4Pb; 0.700 mL, 3.5775 mmol) are added sequentially. The reaction mixture is then allowed to warm gradually to -30°C over 30 min. The reaction is quenched when the temperature reaches -30°C with methanol (10 mL) and saturated aqueous NaHCO3 solution (10 mL). The crude product solution is diluted with ethyl acetate (100 mL) and washed with saturated aqueous NaHCO3 solution (2 x 50 mL) and brine (2 x 50 mL). The organic solution is dried over MgSO4, filtered and concentrated. Purification of the residue by flash chromatography (1:1:2 EtOAc/CH2Cl2/hexane) provides the title compound as a mixture of C7-α-diastereomers.

EXAMPLE 24D 7β-(1-oxopropyl)-17β-t-butyldimethylsilyloxy-4-t-butyldimethylsilyl-4-aza-5α-androstan-3-one

7β-(1-Hydroxypropyl)-17β-t-butyldimethylsiloxy-4-t-butyldimethylsilyl-4-aza-5α- androstan-3-one from Example 24C is oxidized in a manner as in Example 6D to give the title compound.

EXAMPLE 24E 17β-Cyclopropyloxy-7β-(1-oxopropyl)-4-aza-5α-androstan-3-one

7β-(1-Oxopropyl)-17β-t-butyldimethylsilyloxy-4-t-butyldimethylsilyl-4-aza-5α-androstan-3-one from Example 24D is desilylated as in Example 19C and etherified and cyclopropanated as in Preparation 7D to give the title compound. The compound has the following structural formula:

EXAMPLE 25A 17β-Acetoxy-7β-p-tolyl-4-azaandrost-5-en-3-one

Under an inert atmosphere, a slurry of magnesium turnings (2.500 g, 102.8 mmol) in THF (50 mL) is prepared, followed by the addition of 4-bromotoluene (0.50 mL, 4.063 mmol). After the Grignard reaction is initiated, a solution of 4-bromotoluene (10.75 mL, 87.363 mmol) in THF (100 mL) is added dropwise over 60 min while maintaining the reaction temperature below reflux with a water bath. The Grignard solution is stirred for an additional 14 hours at room temperature after addition of the bromide solution, after which a solution of 3β-acetoxy-17β-tert-butyldimethylsilyloxyandrost-5-en-7-one (40.000 g, 86.821 mmol) in THF (500 mL) prepared in Example 16B or obtained otherwise is added while maintaining an inert atmosphere. The addition is allowed to proceed over 24 hours at room temperature. The reaction mixture is then poured into saturated aqueous NH4Cl solution (500 mL), the product extracted into EtOAc (3 x 500 mL); washed with saturated aqueous NaHCO3 solution (3 x 500 mL) and brine (500 mL), dried over MgSO4 and dried over 1 h. dried, filtered and concentrated to afford a crude mixture of 17β-tert-butyldimethylsilyloxy-7-p-tolylandrost-5-ene-7,3β-diol diastereomers (diastereomers).

The diastereomeric mixture prepared above is oxidized as in Example 16D to give 17β-tert-butyldimethylsilyloxy-7β-p-tolylandrosta-4,6-dien-3-one, which is then reduced (hydrogenated) as in Example 16E to give 17β-tert-butyldimethylsilyloxy-7β-p-tolylandrost-5-en-3-one (C₅-olefin). The C₅-olefin is isomerized and desilylated as in Example 16F to give 17β-hydroxy-7β-p-tolylandrost-4-en-3-one which is converted to 17β-hydroxy-5-oxo-7β-p-tolyl-4-nor-3,5-seco-androstane-3-carboxylic acid (seco acid) as in Example 6A. The seco acid is then converted to 17β-acetoxy-7β-p-tolyl-4-azaandrost-5-en-3-one.

EXAMPLE 25B 17β-Cyclopropyloxy-7β-p-tolyl-4-azaandrost-5-en-3-one

17β-Acetoxy-7β-p-tolyl-4-azaandrost-5-en-3-one from Example 25A is hydrolyzed as in Preparation 7C to give the corresponding 17-alcohol and this compound is etherified and cyclopropanated as in Preparation 7D to give the title compound.

EXAMPLE 26A 17β-Hydroxy-7β-p-tolyl-4-aza-5α-androstan-3-one

Hydrogenate 17β-acetoxy-7β-p-tolyl-4-azaandrost-5-en-3-one as in Example 6C to give 17β-acetoxy-7β-p-tolyl-4-aza-5α-androstan-3-one, then hydrolyze as in Preparation 7C to give the title compound.

EXAMPLE 26B 17β-Cyclopropyloxy-7β-p-tolyl-4-aza-5α-androstan-3-one

The 17β-hydroxy-7β-p-tolyl-4-aza-5α-androstan-3-one prepared in Example 26A is etherified and cyclopropanated as in Preparation 7D to give the title compound.

EXAMPLE 26C 17β-Cyclopropylamino-7β-p-tolyl-4-aza-5α-androstan-3-one

The 17β-hydroxy-7β-p-tolyl-4-aza-5α-androstan-3-one prepared in Example 26A is C17 oxidized and cycloaminated as in Examples 6D and 6E, respectively, to give the title compound.

EXAMPLE 27 16β-Propyl-4-methyl-4-azaandrostene EXAMPLE 27A 16β-(2-Propen-1-yl)-4-methyl-4-azaandrost-5-en-3,17-dione

Androst-4-ene-3,17-dione is oxidized to 5-oxo-4-nor-3,5-seco-androstane-5,17-dione (seco acid) as in Example 6A. The seco acid (9.000 g, 31.423 mmol) is slurried with methylammonium chloride (19.5000 g, 288.80 mmol) in acetic acid (75 mL) and heated to reflux under an inert atmosphere. After 3 days, the reaction mixture is allowed to cool to room temperature and diluted with ethyl acetate (EtOAc, 500 mL). The organic solution is washed with water (3 x 200 mL), saturated aqueous NaHCO3 solution (2 x 200 mL) and brine (200 mL), then dried over magnesium sulfate (MgSO4), filtered, concentrated and purified by flash chromatography (1:1:2 EtOAc/CH2Cl2/hexane) to afford 4-methyl-4-azaandrost-5-ene-3,17-dione (azadione). Following the procedure described in N. I. Carruthers et al., J. Org. Chem. 1992, 57(3), 961-965, the azadione prepared above (3.000 g, 9.9529 mmol) is dissolved in CH2Cl2. (50 mL) and diethyl oxalate (1.50 mL, 11.04 mmol) and sodium methoxide (0.750 g, 13.88 mmol) are added sequentially. The reaction mixture is stirred for 60 minutes at 0 °C under nitrogen and a second charge of sodium methoxide (0.100 g, 1.851 mmol) is added. The reaction mixture is stirred for an additional 30 minutes and a third charge of sodium methoxide (0.100 g, 1.851 mmol) and another portion of diethyl oxalate (0.30 mL, 2.209 mmol) are added. The reaction solution is allowed to warm to room temperature and stirred under nitrogen for 16 hours. The solution is then evaporated to dryness and taken up in acetone (50 mL). The acetone solution is transferred to a 100 mL Ace Glass® pressure tube and treated with methyl iodide (3.50 mL, 56.22 mmol). The pressure tube is capped and heated at 55ºC for 22 hours. At the end of this time, the pressure tube is cooled to 0ºC and gently vented. The solvent and reagent are evaporated and the residue is slurried in methanol (50 mL) at 0ºC and sodium methoxide solution (25% in methanol; 2.25 mL, 10.14 mmol) is added to the cold methanolic steroid slurry. After stirring at 0ºC for 90 min, the basic methanol slurry is poured into a 0.5 M acetic acid solution (25.0 mL) with stirring at 0ºC and the resulting precipitate is collected by filtration. The product is purified by flash chromatography to give 4-methyl-16β-(2-propen-1-yl)-4-azaandrost-5-ene-3,17-dione.

EXAMPLE 27B 17β-Cyclopropylamino-4-methyl-16β-(2-propen-1-yl)-4-azaandrost-5-en-3-one

The 4-methyl-16β-(2-propen-1-yl-4-azaandrost-5-ene-3,17-dione prepared in Example 27A or otherwise obtained is cycloaminated as in Example 6E to give the title compound. The compound has the formula:

EXAMPLE 27C 17β-Cyclopropylamino-4-methyl-16β-(2-propen-1-yl)-4-aza-5α-androst-5-en-3-one

17β-Cyclopropylamino-4-methyl-16β-(2-propen-1-yl)-4-azaandrost-5-en-3-one (0.500 g, 1.293 mmol) obtained from Example 27B or otherwise and palladium catalyst (5% Pd on C; 0.100 g, 0.940 mmol) are placed in a 500 mL Parr bottle under nitrogen. Ethanol (100 mL) is added to the reaction vessel and the bottle is charged with H₂ to 60 p.s.i. The hydrogenation reaction is carried out at 60 °C with shaking in a Parr apparatus. After 5 days, the reaction mixture is filtered through Celite® and the filtrate is evaporated to dryness. Purification by flash chromatography (1:1:2 EtOAc/CH2Cl2/hexane) gives the title compound.

EXAMPLE 27D 17β-Cyclopropyloxy-4-methyl-16β-(2-propen-1-yl)-4-azaandrost-5-en-3-one

The 4-methyl-16β-(2-propen-1-yl)-4-azaandrost-5-ene-3,17-dione prepared in Example 27A or otherwise obtained is reduced to give the C17 alcohol as in Example 20A and then etherified and cyclopropanated as in Preparation 7D to give the title compound.

EXAMPLE 27E 17β-Cyclopropyloxy-4-methyl-16β-propyl-4-aza-5α-androstan-3-one

The 17β-cyclopropyloxy-4-methyl-16β-(2-propen-1-yl)-4-azaandrost-5-en-3-one prepared in Example 27D or otherwise obtained is reduced as in Example 27C to give the title compound.

EXAMPLE 28 15β-Ethyl-17β-cyclopropylamino-4-methyl-4-azaandrost-5-en-3-one EXAMPLE 28A 16α-Bromo-17,17-ethylenedioxyandrost-5-en-3β-ol-3-acetate

3β-Acetoxyandrost-5-en-17-one (dehydroisoandrosterone-3-acetate, ALDRICH 39,008-9) is ketalized as described in Example 18A to give 17,17-ethylenedioxyandrost-5-en-3β-ol-3-acetate (C₁₇-ketal). The C₁₇-ketal is dissolved in a solution of dry THF (150 mL) and a solution of pyridinium bromide perbromide (ALDRICH 21,469-8, C₅H₅NH⁺Br₃⁻; 88.000 g, 275.138 mmol) in dry THF (150 mL) is added under an inert atmosphere. The reaction mixture is stirred at ambient temperature for 2 hours, then treated with NaI (70.000 g, 467.01 mmol). The reaction solution is stirred for an additional 30 minutes, then treated with a mixture of sodium thiosulfate (Na2S2O3, 95.000 g, 600.85 mmol) in a mixture of water (150 mL) and pyridine (35.0 mL, 432.7 mmol). The resulting solution is stirred at room temperature for an additional 3 hours. The reaction mixture is then diluted with water (300 mL) and THF is evaporated under reduced pressure. The precipitate that forms is collected by filtration and recrystallized from ethanol to give the title compound.

EXAMPLE 28B 17,17-Ethylenedioxyandrost-5,15(16)-dien-3β-ol acetate

16α-Bromo-17,17-ethylenedioxyandrost-5-en-3β-ol-3-acetate from Example 28A (50.000 g, 110.28 mmol) and potassium t-butoxide are dissolved in dimethyl sulfoxide (DMSO; 500 mL) and the solution is warmed to 45 °C under an inert atmosphere. After 22 hours, the reaction mixture is partitioned between ether (2000 mL) and water (750 mL). The layers are separated and the ether solution is washed with brine (2 x 500 mL), dried over MgSO4, filtered and evaporated to dryness. The residue is recrystallized from ethanol-water to give the title compound.

EXAMPLE 28C 3β-t-Butyldimethylsilyloxyandrost-5,15(16)-dien-17-one

17,17-Ethylenedioxyandrost-5,15(16)-dien-3β-ol acetate from Example 28B or otherwise obtained is dissolved in acetone (500 mL) and water (60 mL) and the solution is cooled to 0ºC. To the solution at 0ºC is added p-toluenesulfonic acid monohydrate (1.000 g, 5.257 mmol). The reaction mixture at 0ºC is stirred for 5 hours and then stored at 4ºC for 16 hours. The quenched reaction solution is diluted with water (300 mL) and acetone is evaporated under reduced pressure. The remaining aqueous slurry is extracted with EtOAc (2 x 300 mL) and the combined organic layers are washed with saturated aqueous NaHCO3 solution (200 mL) and brine (200 mL), dried over MgSO4, filtered and concentrated. The residue is purified by flash chromatography (1:1 EtOAc/CH2Cl2) to afford 3-hydroxyandrost-5,15(16)-dien-17-one (C3 alcohol).

The C3 alcohol prepared above is silylated as in Example 16A to give the title compound.

EXAMPLE 28D 15β-Ethylandrost-4-en-3,17-dione

Copper(I) chloride (CuCl, 0.7611 g, 7.689 mmol) was slurried in THF (40 mL) under nitrogen and then quenched to -22 °C with a dry ice/tetrachloroethylene bath. Ethylmagnesium chloride solution (2.00 M in ether; 22.0 mL, 44.0 mmol) was added to the cold copper(I) chloride slurry and the dark solution was stirred for 90 min. A solution of 3β-t-butyldimethylsilyloxyandrost-5,15(16)-dien-17-one (from Example 28C) in THF (50 mL) was added dropwise to the organometallic solution via polyethylene cannula over 10 min, followed by a THF rinse (20 mL). The reaction mixture is stirred at -22°C for 2 hours and then allowed to warm to ambient temperature over 30 minutes, followed by addition of saturated aqueous NH4Cl solution (50 mL). The biphasic mixture is extracted with EtOAc (2 x 40 mL) and the combined organic layers are washed with brine (50 mL), dried over MgSO4, filtered and evaporated to dryness. Purification by flash chromatography gives 15β-ethyl-3β-t-butyldimethylsilyloxyandrost-5-en-17-one, which is deprotected as in Example 19C to give the corresponding 3-alcohol which is then oxidized as in Example 16D to give the title compound which has the following formula:

EXAMPLE 28E 15β-Ethyl-4-azaandrost-5-en-3,17-dione

15β-Ethylandrost-4-ene-3,17-dione from Example 28D or otherwise obtained is oxidized as in Example 6A to 15β-ethyl-5,17-dioxo-4-nor-3,5-seco-androstane-3-carboxylic acid (seco acid) which is then converted to 15β-ethyl-4-azaandrost-5-ene-3,17-dione as in Example 6B.

EXAMPLE 28F 15β-Ethyl-17β-cyclopropylamino-4-azaandrost-5-en-3-one

15β-Ethyl-4-azaandrost-5-ene-3,17-dione is cycloaminated as in Example 6E to give the title compound.

EXAMPLE 29 15β-Ethyl-17β-cyclopropyloxy-4-azaandrost-5-en-3-one EXAMPLE 29A 15β-Ethyl-17β-hydroxy-4-azaandrost-5-en-3-one

15β-Ethyl-4-azaandrost-5-ene-3,17-dione is reduced as in Example 20A to give the title compound.

EXAMPLE 29B 15β-Ethyl-17β-cyclopropyloxy-4-azaandrost-5-en-3-one

15β-Ethyl-17β-hydroxy-4-azaandrost-5-en-3-one from Example 29A is etherified and cyclopropanated as in Preparation 7D to give the title compound.

EXAMPLE 29C 15β-Ethyl-17β-cyclopropyloxy-4-aza-5α-androst-3-one

15β-Ethyl-17β-hydroxy-4-azaandrost-5-en-3-one is hydrogenated as in Example 27C and then etherified and cyclopropanated as in Preparation 7D to give the title compound.

Claims (30)

1. Compound of the following general formula: in the: A means O or NH; R¹ represents H or C₁₋₄alkyl; R² represents H, a halogen atom, a phenylthio, phenylsulfinyl or phenylsulfonyl group ; R³ represents H, a halogen atom, a C₁₋₄alkylthio, C₁₋₄alkylsulfinyl or C₁₋₄alkylsulfonyl radical; R⁴ is H, C₁₋₄ alkyl or C₂₋₄ alkenyl; R⁵ represents H or C₁₋₄ alkyl; A: a) an oxo group; b) (H)(H) or an α-hydrogen atom and a β-substituent selected from a C₁₋₄alkyl, C₂₋₄alkanyl, a hydroxy group, a C₁₋₄alkanoyloxy, C₁₋₄alkoxycarbonylmethyl, a carboxymethyl group, a C₁₋₄alkoxycarbonyl group, a carboxy group, a C₁₋₄alkanoyl group and a halogen atom; and the possible presence of a double bond means provided that: a) a 1,2-double bond is present when R² is present and is different from a hydrogen atom, b) no 6,7-double bond is present when Z is an oxo group, and c) A does not represent an NH group when R¹ represents a CH₃ group and R², R³, R⁴ and R⁵ are H and Z is (H)(H) and there are no double bonds in any of the positions 1(2), 5(6) or 6(7), or a pharmaceutically acceptable salt thereof. 2. A compound according to claim 1 having the following general formula: in the: A means O or NH; R¹ represents H or C₁₋₄alkyl; R² represents H or a halogen atom; R³ represents H or a halogen atom; and Z represents (H)(H) or an α-hydrogen atom and a β-substituent selected from a C₁₋₄alkyl, C₁₋₄alkoxycarbonylmethyl radical and a carboxymethyl group , or a pharmaceutically acceptable salt thereof. 3. A compound according to claim 2 having the following general formula: in the: A means O or NH; R¹ represents H or C₁₋₄alkyl; R² represents H or a fluorine atom, or a pharmaceutically acceptable salt thereof. 4. A compound according to claim 3 having the following formula: or a pharmaceutically acceptable salt thereof. 5. A compound according to claim 3 having the following formula: or a pharmaceutically acceptable salt thereof. 6. A compound according to claim 3 having the following formula: or a pharmaceutically acceptable salt thereof. 7. A compound according to claim 3 having the following formula: or a pharmaceutically acceptable salt thereof. 8. A compound according to claim 3 having the following formula: or a pharmaceutically acceptable salt thereof. 9. Compound of the formula: in the: A means O or NH; R¹ represents H or C₁₋₄alkyl; R² represents H, a halogen atom, a phenylthio, phenylsulfinyl or phenylsulfonyl group ; R³ is H, a halogen atom, a C₁₋₄-alkylthio, C₁₋₄-alkylsulfinyl or C₁₋₄-alkylsulfonyl radical represents; R⁴ is H, C₁₋₄ alkyl or C₂₋₄ alkenyl; R⁵ represents H or C₁₋₄ alkyl; A: a) an oxo group; b) (H)(H) or an α-hydrogen atom and a β-substituent selected from a C₁₋₄alkyl, C₂₋₄alkenyl radical, a hydroxy group, a C₁₋₄alkanoyloxy, C₁₋₄alkoxycarbonylmethyl radical, a carboxymethyl group, a C₁₋₄alkoxycarbonyl radical, a carboxy group, a C₁₋₄alkanoyl radical and a halogen atom; the possible presence of a double bond means provided that: a) a 1,2-double bond is present when R² is present and is different from a hydrogen atom, and b) no 6,7-double bond is present when Z represents an oxo group, or a pharmaceutically acceptable salt thereof for use as a medicament. 10. A compound according to claim 9 of the formula: in the: A means O or Nile; R¹ represents H or C₁₋₄alkyl; R² represents H or a halogen atom; R³ represents H or a halogen atom; and Z represents (H)(H) or an α-hydrogen atom and a β-substituent selected from a C₁₋₄alkyl, C₁₋₄alkoxycarbonylmethyl radical and a carboxymethyl group , or a pharmaceutically acceptable salt thereof, for use as a medicament. 11. A compound according to claim 9 of the formula: in the: A means O or NH; R¹ represents H or a C₁₋₄ alkyl group; R² represents H or a fluorine atom, or a pharmaceutically acceptable salt thereof, for use as a medicament. 12. Use of a compound according to any one of claims 1 to 11 for manufacturing a medicament for inhibiting C₁₇₋₂₀-lyase and 5α-reductase in a patient in need thereof. 13. Use of a compound according to any one of claims 1 to 5 and 7 to 11 for the manufacture of a medicament for inhibiting C₁₇₋₂₀-lyase in a patient in need thereof. 14. Use according to claim 13, wherein the compound has the formula: having. 15. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for inhibiting 5α-reductase in a patient in need thereof. 16. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for treating androgen dependent disorders in a patient in need thereof. 17. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for treating polycystic ovarian disease in a patient in need thereof. 18. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for treating benign prostatic hyperplasia in a patient in need thereof. 19. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for treating prostate cancer in a patient in need thereof. 20. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for treating a DHT-mediated disease in a patient in need thereof. 21. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for treating acne in a patient in need thereof. 22. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for treating an estrogen-mediated or estrogen-dependent disorder in a patient in need thereof. 23. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for treating breast cancer in a patient in need thereof. 24. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for inhibiting C₁₇-hydroxylase in a patient in need thereof. 25. Use of a compound according to any one of claims 1 to 5 and 9 to 11 for the manufacture of a medicament for treating Cushing's syndrome in a patient in need thereof. 26. A pharmaceutical composition comprising a compound according to any one of claims 1 to 11 in admixture with a pharmaceutically acceptable carrier. 27. A product containing a compound according to any one of claims 1 to 11 and an androgen receptor antagonist as a preparation for combined therapy for the treatment of androgen-dependent disorders. 28. The product of claim 27, wherein the androgen receptor antagonist is flutamide. 29. A product containing a compound according to any one of claims 1 to 11 and an androgen receptor antagonist as a preparation for combined therapy for the treatment of prostate cancer. 30. The product of claim 29, wherein the androgen receptor antagonist is flutamide.